Immunomodulatory peptides of vespid antigen 5

ABSTRACT

The present invention is directed to immunogenic peptides from vespid antigen 5. These immunogenic peptides can be used in immunotherapy for vespid venom allergic individuals. The present invention is thus directed to T cell epitopes of vespid antigen 5 that can anergize T cell responses in sensitive individuals.

The research leading to the present invention was supported by UnitedStates Public Health Service Grant No. AI-17021. The government may havecertain rights in the invention.

This Application is a Division of application Ser. No. 08/614,935 filedMar. 11, 1996 now U.S. Pat. No. 5,804,201, issued Sep. 8, 1998.

FIELD OF THE INVENTION

The present invention is directed to immunogenic peptides from vespidantigen 5. These immunogenic peptides can be used in immunotherapy forvespid venom allergic individuals. The present invention is thusdirected to T cell epitopes of vespid antigen 5 that can anergize T cellresponses in sensitive individuals.

BACKGROUND OF THE INVENTION Biochemical Aspects of Insect VenomAllergens

Insect sting allergy to bees and vespids is of common occurrence. Thevespids include hornets, yellowjackets and wasps [Golden, et al., Am.Med. Assoc., 262:240 (1989)]. Susceptible people can be sensitized onexposure to minute amounts of venom proteins; as little as 2-10 μg ofprotein is injected into the skin on a single sting by a vespid [Hoffmanand Jacobson, Ann. Allergy., 52:276(1984)].

Indeed, venom allergens from insects of the Hymenoptera order have beenextensively studied. These insects are bees, vespids and fire ants. Thebees include honey bees (Apis melifera) and bumble bees (Bombuspennsylvanicua). The vespids include hornets (Dolichovespula spp.; Vespacrabo), yellow jackets (Vespula spp.), and paper wasps (Polistes spp.).In Table 1 are listed the venom allergens from these insects with knownprimary structures.

                  TABLE 1                                                         ______________________________________                                        Cloned and/or sequenced insect venom allergens                                Allergen              Mol.     Recombinant protein.sup.3                      name.sup.1                                                                              Common name Size.sup.2                                                                             unfolded                                                                             folded                                  ______________________________________                                        Bumble bee, Bombus pennsylvanicus                                             Bom p 1   phospholipase A.sub.2                                                                     16 kd    -      -                                       Bom p 4   protease    28 kd    -      -                                       Honey bee, Apis melifera                                                      Api m 1   phospholipase A.sub.2                                                                     16 kd    +      +                                       Api m 2   hyaluronidase                                                                             39 kd    +      +                                       Api m 3   acid phosphatase                                                                          43 kd    -      -                                       Api m 4   melittin     3 kd    -      -                                       Fire ant, Solenopsis invicta                                                  Sol i 1   phospholipase A.sub.1                                                                     37 kd    -      -                                       Sol i 2               30 kd    -      -                                       Sol i 3   antigen 5   23 kd    -      +                                       Sol i 4               20 kd    -      -                                       White face hornet, Dolichovespula maculata                                    Dol m 1   phospholipase A.sub.1                                                                     34 kd    +      -                                       Dol m 2   hyaluronidase                                                                             38 kd    +      -                                       Dol m 5   antigen 5   23 kd    +      +                                       European hornet, Vespa crabo                                                  Vesp c 1  phospholipase A.sub.1                                                                     23 kd    -      -                                       Vesp c 5  antigen 5   23 kd    -      -                                       Paper wasp, Polistes annularis                                                Pol 1 5.sup.5                                                                           antigen 5   23 kd    +      -                                       Yellow jacket, Vepula vulgaris                                                Ves v 1   phospholipase A.sub.1                                                                     34 kd    +      -                                       Ves v 2   hyaluronidase                                                                             38 kd    +      -                                       Ves v 5.sup.5                                                                           antigen 5   23 kd    +      -                                       ______________________________________                                         Footnotes                                                                     .sup.1 Allergen names are designated according to an accepted nomenclatur     system [King et al., WHO Bulletin, 72:797 (1994)].                            .sup.2 Several allergens are glycoproteins, and the molecular size given      refers only to the protein portion.                                           .sup.3 + and - signs refer to the availability of recombinant proteins.       .sup.4 Sequence of antigen 5 from S. richteria is known [Smith & Hoffman,     J. Allerg. Clin. Imunol., 89:293 (1992)].                                     .sup.5 Sequences of antigen 5s from several other vespids are known; D.       arenaria [Lu et al., J. Immunol. 150:2823 (1993)], P. exclamans and P.        fuscatus, and V. flavopilosa, V. germanica, V. maculifrons, V.                pennsylvanica, V. spamosa and V. vidua [Hoffman et al., Int. Archs.           Allergy App. Immunol., 84:24 (1987)].                                    

There are many species of hornets (genus Dolichovespula), yellowjackets(genus Vespula) and wasp (genus Polistes) in North America [Akre, etal., "Yellowjackets of America North of Mexico," Agriculture Hand bookNo. 552, US Department of Agriculture (1980)]. The vespids have similarvenom compositions [King, et al., Biochemistry, 17:5165 (1978); King, etal., Mol. Immunol. 20:297 (1983); King, et al., Arch. Biochem. Biophys.230:1 (1984); King, et al., J. Allergy and Clin. Immunol., 75:621(1985); King, J. Allergy Clin. Immunol., 79:113 (1987); Hoffman, J.Allergy and Clin. Immunol., 75:611 (1985)]. Their venom each containsthree major venom allergens, phospholipase (37 kD), hyaluronidase (43kD) and antigen 5 (23 kD) of as yet unklnown biologic function.Homologous venom allergens from hornets, yellow jackets, and paper waspshave high degrees of sequence identity ranging form about 70% forantigen 5s to about 90% for hyaluronidases [Lu et al., J. Immunol.,150:2823 (1993)].

Antigen 5 from several species each of hornets, yellowjackets and paperwasps have been cloned and/or sequenced [Fang et al., Proc. Natl. Acad.Sci., USA, 85:895-899 (1988); Lu et al., supra; Hoffman, J. AllergyClin. Immunol., 92:707-716 (1993)]. For phospholipases andhyaluronidases only those from hornets and yellowjackets have beencloned and/or sequenced [Soladatova et al., FEBS Letters, 320:145-149(1993); Lu et al., J. Biol. Chem., 270 :4457-4465 (1995); King et al.,J. Allergy Clin. Immunol., In press (1996); Hoffman, Int. Arch. AllergyImmunol., 104:184-190 (1994)]. One common feature of these venomproteins is their varying extents of sequence homology with mammalianproteins.

White faced hornet (Dolichovespula maculata) has three forms of antigen5. Two of these forms, Dol m 5.01 and 5.02, differ in 23% of theirsequences, and they are antigenically cross reactive at both B and Tcell levels [Fang et al., Proc. Natl. Acad. Sci., USA, 85:895-899(1988); Lu et al., J. Immunol., 150:2823-2830 (1993)]. The studiesdescribed here were made with Dol m 5.02, also referred to as hornet Ag5form 2. The amino acid sequences of Dol m 5.01 and 5.02, as well asthose of the homologous antigen 5s from yellow hornet (Dolichovespulaarenaria), two species of yellowjacket (Vespula maculifrons andvulgaris) and two species of papers wasps (Polistes annularis andexcoamans) are given in FIG. 1.

Fire ant venom contains four allergens. They are antigen 5,phospholipase A₁, and Sol i 2 and 4. Fire ant antigen 5 has about 50%sequence identity with vespid antigen 5 [Hoffman, J. Allergy Clin.Immunol., 91:71 (1995)]. Only partial sequence data is available forfire ant phospholipase, and it shows sequence identity with vespidphospholipase A₁ [Hoffman, J. Allergy Clin. Immunol., 95:372 (1995)].

Bumble bee venom has two allergens of known sequences; phospholipase A₂and protease. But honey bee venom has four allergens of know sequences;acid phosphatase, phospholipase A₂, hyaluronidase and a cytolyticpeptide melittin. The two bee venom phospholipase A₂ have extensivesequence identity and they are not-related to vespid phospholipase A₁[Hoffman, "Hymenoptia Venom Proteins" in National Toxins, R. B. Singh(ed.), Plenum Publishing 6 (1996)]. Honey bee venom hyaluronidase hasabout 55% sequence identity with the homologous vespid hyaluronidases.

In addition to the insect venom allergens described above, the completeamino acid sequence of several major allergens from different grass[Perez, et al., J. Biol. Chem., 265:16210 (1990); Ansari, et al.,Biochemistry, 26:8665 (1989); Silvanovich, et al., J. Biol. Chem.,266:1204 (1991)], tree pollen [Breiteneder, EMBO J., 8:1935(1989);Valenta, et al., Science, 253:557 (1991)], weed pollen [Rafnar, et al.,J. Biol. Chem., 266:1229 (1991); Griffith, et al., Int. Arch. AllergyAppl. Immunol., 96:296 (1991)], mites (Chua, et al., J. Exp. Med.,167:175 (1988)], cat dander [Griffith, et al., Gene., 113:263 (1992)],and mold [Aruda, et al., J. Exp. Med., 172:1529 (1990); Han, et al., J.Allergy Clin. Immunol., 87:327 (1991)] have been reported in the pastfew years. These major allergens are proteins of 10-40 kD and they havewidely different biological functions. Nearly all allergens of knownsequences have a varying extent of sequence similarity with otherproteins in our environment.

T and B Cell Epitopes of Allergens

Antibody responses to proteins require the collaboration of T helper andB lymphocytes and antigen presenting cells (APC). The antigen receptorsof B cells are the membrane-bound antibody (Ab) molecules, whichrecognize and bind immunogens directly. The antigen receptors of T cells(TCR) only recognize and bind complexes of antigenic peptide-MHC classII molecule. Immunogens are first processed by APC into peptides thatare presented on the surface of APC in association with the MHC class IImolecules [Unanue, Current Opinion in Immunol, 4:63 (1992)]. As MHCmolecules are highly polymorphic in individuals, they have differentspecificity of binding antigenic peptides [Rothbard and Gefter, Ann.Rev. Immunol., 9:527 (1991)]. This is one mechanism for genetic controlof immune response.

T helper cells are activated when the antigen receptor binds thepeptide-MHC complex on the surface of APC. Activated T cells secretelymphokines. In mice [Street and Mosmann, FASEB J., 5:171 (1991)] andapparently in humans [Wierenga. et at., J. Immunol., 144:4651 (1990);Parronchi, et al., Proc. Natl. Acad. Sci. USA., 88:4538 (1991)] the Thelper cells can be divided into different types on the basis of theirpatterns of lymphokine production. Primarily, T helper cells divide intotwo groups: TH1 cells producing IL-2 and IFN-γ, and TH2 cells producingIL-4 and IL-5. These lymphokines in turn influence the antigen-activatedB cells to differentiate and proliferate into plasma cells secreting Absof different isotypes. IL-4 is one lymphokine known to influence IgEsynthesis [Finkelman. et al., Ann. Rev. Immunol., 8:303 (1990)].

It is believed that the entire accessible surface of a protein moleculecan be recognized as epitopes by the antigen receptors of B cells,although all epitopes are not necessarily recognized with equallikelihood [Benjamin, et al., Ann. Rev. Immunol., 2:67 (1984)]. B cellepitopes of a protein are of two types: topographic and linear. Thetopographic type consists of amino acid residues which are spatiallyadjacent but may or may not be sequentially adjacent. The linear typeconsists of only sequentially adjacent residues. X-ray crystallographicdata of Ag-Ab complexes indicate the size of their complementary bindingregion to have 16-17 amino acid residues [Amit, et al., Science, 233:747(1986)], but peptide mapping suggests that less than about 8 residuescontribute significantly to the binding process of a linear epitope[Appel, et al., J. Immunol., 144:976 (1990)].

Allergens, like other protein antigens, can have both types of B cellepitopes or only one. For example, vespid antigen 5s have both types[King et al., J. Immunol., 154:577 (1995)]. Bee venom melittin appearsto have only one B cell epitope of linear type [King, et al., J.Immunol., 133:2668 (1984)].

T cell epitopes of proteins consist of only the linear type since theyare peptides that have been processed in the lysosomes of APC byproteases of unknown specificity [Unanue, Curr. Op. Immunol., 4:63(1992)]. Analysis of naturally processed antigenic peptides bound to MHCclass II molecules indicates that their size ranges from about 13 to 17amino acid residues, but analysis of synthetic peptide-MHC class IImolecule complex for their T cell proliferate response suggests aminimal size of about 8 amino acid residues [Cf. Rudensky et al.,Nature, 353:622 (1991)]. Studies suggest that T cell epitopes aredistributed throughout the entire protein molecule, and they mayfunction as major or minor determinants depending on the MHC haplotypeof the immunized host [Roy, et al., Science, 244:572; Gammon, et al.,Immunol. Rev., 98:53 (1987); O'Hehir et al., Ann. Rev. Immunol., 9:67(1991)].

Hypersensitivity of the immediate type is known to be caused by thepresence of allergen-specific IgE. IgE is found in the circulation andbound to specific IgE-Fc receptors on mast cells and basophils.Cross-linking of cell-bound IgE by allergens leads to release ofhistamine, leukotrienes and other chemical mediators that cause theallergic symptoms. IgE is one of the different isotypes ofimmunoglobulins. As pointed out above, lymphokines secreted by T cellsinfluence isotype switch events in B cells.

Because of the central role of TH2 cells in determining the isotypesswitch event of B cells, the T cell epitopes of several allergens havebeen mapped [Cf. O'Hehir et al., supra]. These allergens include ragweedAmb a III, rye grass Lol p I, cat Fel d I, mouse urine Mus m I, midgeChi t I, bee venom phospholipase A₂ [Dhillon, et al., J. Allergy Clin.Immunol., 90:42 (1992)] melittin [Fehlner, et al., J. Immunol., 146:799(1991)], and hornet antigen 5 [King et al., J. Allergy Clin. Immunol.,91:283 (1993)]. The data do not reveal any unusual or common structuralfeatures. However, any conclusion from these data is qualified as thesedata are collected from humans and mice of different haplotypes.

Modulation of T and B Cell Responses

Normally hosts are tolerant to the dominant B and T cell epitopes ofself proteins by clonal deletion and anergy. However this tolerance canbe broken under certain circumstances [Gammon, et al., Immunol. Today.,12: 93 (1991); Basten, et al., Immunol. Rev., 122:5 (1991)]. It has beensuggested that self-tolerance is broken in autoimmune diseases throughencounters with foreign proteins that are similar to host proteins.Therefore the sequence similarity of allergens with autologous proteinsis of interest for closer investigation.

Mature B cells are activated in response to multi-valent antigens whichcan cross-link cell surface Ig receptors [DeFranco. Ann. Rev. CellBiol., 3:143 (1987)], and they are rendered anergic in response tomono-valent antigen [Basten, et al., 1991, supra]. Antigen activation ofT cells requires not only the integration of TCR with peptide-MHCcomplex but also with other co-stimulating signals on the surface of APC[Schwartz, Science, 248:1349 (1990); Jenkins and Miller, FASEB J.,6:2428 (1992)]. Interaction of TCR with peptide-MHC complex in absenceof co-stimulating signals can lead to T cell anergy.

The molecular mechanism of B or T cell anergy is not yet understood [Cf.Schwartz, 1990, supra; Jenkins and Miller, 1992, supra; Ales-Martinez,et al., Immunol. Today, 12:201 (1991)]. In vitro studies with T cellclones reveals that occupancy of TCR by artificial peptide-MHC complexin absence of co-stimulating signals leads to altered intracellularsignal transduction and/or repressor gene activation which can preventlymphokine transcription.

Early studies have shown that the physical state of the immunogen andthe route of immunization are important variables in determining theoutcome of an immune response. In the light of our currentunderstanding, these variables may well influence antigen presentationso as to have T and B cell activation or anergy.

One way to treat allergic diseases is by immunotherapy which involvesrepeated subcutaneous injections of the offending allergen(s) intopatients. The amounts of allergens which can be injected are limited bythe danger of unwanted systemic allergic reaction in patients. For mostpatients following immunotherapy, their allergen-specific IgE levelsinitially rise followed with gradual decrease of their allergen-specificIgE levels, and there is also downregulation of allergen-specific T cellresponses [P. S. Norman, Current Op. Immunol., 5:968 (1993)].

Because of the undesirable systemic reaction on immunotherapy withnative allergens, there has been continued interest in the developmentof modified allergens with reduced allergenic activities forimrnmunotherapy [T. P. King. in "Bronchial Asthma," edited by E. B.Weiss and M. Stein, Little Brown, Boston, pp. 43-49 (1993); R.E. O'Hehiret al., 1991, supra].

Three reports have appeared recently on the use of T cell epitopepeptides to modulate allergen-specific immune responses. One report ison the subcutaneous injection of mice with two peptides from the majorcat allergen Fel d I to decrease T cell response to the entire moleculeFel d I [Briner et al., Proc. Natl. Acad. Sci. U.S.A., 90:7608-12(1993)]. Another is on the intranasal therapy with a peptide from themajor mite allergen Der p I to suppress allergen-specific response innaive or sensitized mice [Hoyne et al., J. Exp. Med., 178:1783-1788(1993)]. The third reports that peptides containing T cell epitopes ofbee venom phospholypase A₂ were used successfully in immunotherapy totreat patients with bee venom allergy [Muller et al., J. Allergy Clin.Immunol. 97:426 (1996)].

Since an MHC class II molecule of any one haplotype can bind a widerange of peptides in its binding groove, it may Pe possible to modulateT cell response by inhibition of allergen-derived T cell epitope bindingto MHC molecules with other peptides. For example, a mouse lysozymepeptide which is not immunogenic by itself in H-2^(k) mice inhibits Tcell response to hen egg white lysozyme [Adorini and Nagy, Immunol.Today., 11:21 (1990)]. Another example is the in vitro inhibition of Tcell response to a mite allergen by an influenza HA peptide [O'Hehir etal., J. Allergy Clin. Immunol., 87:1120 (1991)].

Experimental autoimmune encephalomyelitis (EAE) in mice or rats is awell-studied model for multiple sclerosis. Many studies have identifiedimmunodominant T cell determinants for myelin basic protein, which isused to induce this condition. Peptides that correspond toimmunodominant epitopes of myelin basic protein can induce tolerance tothe same peptide antigen or to the intact myelin basic protein. The samepeptides that induced tolerance could also induce T cell anergy in anongoing autoinmmune response [Gaur et al., Science, 259:1491-1494(1992)].

It has been reported that subcutaneous or intranasal pretreatment ofmice with T cell epitopes peptides of the major cat allergen Fel d 1[Briner et al., Proc. Natl. Acad. Sci. USA, 90:7608-7612 (1993)] or themajor mite allergen Der p 1 [Hoyne et al., J. Exp. Med., 178:1783-1788(1993)] lead to T cell anergy and that reduced antibody responses areobserved on subsequent immunization with the allergen. T cell epitopepeptides of Fel d 1 studied in the murine system are currently beingevaluated for immunotherapy of patients [Norman et al., J. Allergy Clin.Immunol., 95:259 (1995)]. Published data indicate that the T cellepitope regions of Der p 1 detected in the murine system overlap thosefound in humans [O'Hehir et al., Eruop. J. Clin. Invest., 23:763(1993)]. On the basis of findings with Fel d 1 and Der p 1, it isreasonable to conclude that the T cell epitope data of white facedhornet Ag5 obtained in mice as described herein will be applicable forstudies in humans.

There remains a need in the art for the identification of T cellepitopes of vespid venom allergens, particularly antigen 5, forimmunotherapy of venom allergy.

There is also a need in the art to use peptides having T cell epitopesof vespid venom allergens to study induction of tolerance in mice andinduction of tolerance in humans.

There is a further need to test whether a modified peptide inhibitsallergen T cell epitope binding to MHC class II molecule, or induces Tcell anergy, or both.

These and other needs in the art are satisfied by the present invention.

The citation of references herein shall not be construed as an admissionthat such is prior art to the present invention.

SUMMARY OF THE INVENTION

The present invention provides the sequences of immunodominant peptidesof vespid venom antigen 5, and corresponding peptides from otherantigens, such as fire ant Sol i 3. An immunodominant peptide is onethat contains a T cell epitope of the antigen, such that T cellsimmunized with the antigen will be stimulated when contacted with thepeptide. Such peptides of the invention are preferably immunomodulatorypeptides as well, in that they induce T cell anergy when administered toa subject, or otherwise affect the immune response of the subject.

In yet another embodiment, the present invention provides apharmaceutical composition effective for the treatment of a vespid venomallergen-specific allergic condition comprising a polypeptide of theinvention that has an immunomodulatory portion of a T cell epitope of avespid venom antigen 5. More particularly, the invention providespharmaceutical compositions comprising such polypeptides, includingdifferent isoforms, of a vespid venom antigen 5, for example,Dolichovespula maculata, Vespula vulgaris, Vespula maculifrons,Dolichovespula arenaria, Polistes annularis, and Polistes exclamans.

In yet still another embodiment, the present invention provides a methodfor treating a vespid venom allergen-specific condition comprisingadministering a therapeutically effective dose of a pharmaceuticalcomposition of the invention.

In its broadest aspect, the present invention is directed to a peptidecharacterized by having between 8 (the generally recognized minimumnumber of amino acids for an antigenic T cell epitope) and 35 amino acidresidues of vespid venom antigen 5; and being antigenic for T cellproliferation in a mouse immunized with a vespid venom antigen 5, whichmouse is a strain selected from the group consisting or BALB/c, ASW/Sn,C3H/He, and P/J. More particularly, a peptide of the inventioncorresponds to a fragment of white face hornet antigen 5, form 2,selected from the group consisting of:NNYCKIKCRKGIHTLCKFGT; (SEQ IDNO:8)GIHTLCKFGTSMKPNCGRNV; (SEQ ID NO:9)KNEILKRHNDFRQNVAKGLE; (SEQ IDNO:12)FRQNVAKGLETRGKPGPQPP; (SEQ ID NO:13)TRGKPGPQPPAKNMNVLVWN; (SEQ IDNO:14)DELAKIAQTWANQCDFNHDD; (SEQ ID NO:16)CRNTAKYQVGQNIAISSTTA; (SEQ IDNO:18)QNIAISSTTATQFDRPSKLI; (SEQ ID NO:19)TQFDRPSKLIKQWEDEVTEF; (SEQ IDNO:20)KQWEDEVTEFNYKVGLQNSN; (SEQ ID NO:21)NYKVGLQNSNFRKVGHYTQM; (SEQ IDNO:22)FRKVGHYTQMVWGKT; (SEQ ID NO:23)KEIGCGSIKYIEDNWYTHYL; and (SEQ IDNO:25)IEDNWYTHYLVCNYGPGGND. (SEQ ID NO:26)

Such peptides may be from the specific antigen, white face hornetantigen 5, form 2, or may be from the corresponding segments of othervespid venom antigens, such as but not limited to Dolichovespulamaculata, Vespula vulgaris, Vespula maculifrons, Dolichovespulaarenaria, and Polistes exclamans. Other sources of antigen 5 of theinvention include Vespa crabo (European hornet), V. flavopilosa (yellowjacket), V. germanica (yellowjacket), V. pennsylvannica (yellowjacket),V. squamosa (yellowjacket), V. vidue (yellowjacket), and P fuscatus(paperwasp), or fire ant allergen Sol i 3, which shares sequencesimilarity with (and possibly is homologous to) vespid venom antigen 5s.Such corresponding peptides from various species are referred to hereinas homologs, as they represent variants from the corresponding, i.e.,homologous, alleles of the antigen 5 gene from different vespid species.The term allelic variant includes, as well, single amino acid variationsin antigen 5s from the same species, including different isoforms (suchas the three isoforms found for white face hornet antigen 5), andmutations.

Preferably, the peptide corresponds to a fragment of white face hornetantigen 5, form 2, selected from the group consisting of:

    NNYCKIKCRKGIHTLCKFGT;                                                                             (SEQ ID NO:8)                                             GIHTLCKFGTSMKPNCGRNV;                                                                             (SEQ ID NO:9)                                             KNEILKRHNDFRQNVAKGLE;                                                                             (SEQ ID NO:12)                                            FRQNVAKGLETRGKPGPQPP;                                                                             (SEQ ID NO:13)                                            DELAKIAQTWANQCDFNHDD;                                                                             (SEQ ID NO:16)                                            NYKVGLQNSNFRKVGHYTQM;                                                                             (SEQ ID NO:22)                                            FRKVGHYTQMVWGKT; and                                                                              (SEQ ID NO:23)                                            IEDNWYTHYLVCNYGPGGND.                                                                             (SEQ ID NO:26)                                        

This group of peptides showed significant T cell stimulation of antigen5-immunized T cells in more than one species.

As pointed out above, the present invention advantageously provides notonly the immunodominant T cell epitopes for white face hornet antigen 5,but corresponding epitopes from antigen 5s from other vespid species,particularly subfamilies Vespinae and Polistinae, more particularly thegenera Vespa, Vespula, Dolichovespula, and Polistes. In addition, theinvention provides for consensus polypeptides, which incorporate thesame amino acid residues at conserved positions, and any amino acidresidue or amino acid residues having similar properties at divergentpositions. In other words, the present invention contemplatesimmunodominant peptides that incorporate polymorphisms found among theantigen 5 homologs from different species.

In a specific embodiment, the peptide has the amino acid sequence X₁BYCKIX₂ CX₃ X₄ GX₅ X₆ HTX₇ CX₈ X₉ G (SEQ ID NO:31), wherein X₁ is aneutral amino acid residue, X₂ is a basic amino acid residue or deleted,X₃ is any amino acid residue, X₄ is a polar amino acid, X₅ is glycine ordeleted, X₆ is any amino acid residue, X₇ is an amino acid residue withan aliphatic side chain, X₈ is a polar amino acid residue, and X₉ is anamino acid residue with an aromatic side chain. In a more specificembodiment, X₁ is valine or asparagine; X₂ is lysine, arginine, ordeleted; X₃ is proline, arginine, serine, or leucine; X₄ is arginine,lysine, or serine; X₆ isoleucine, leucine, threonine, or valine; X₇ isalanine, valine, isoleucine, or leucine; X₈ is lysine, arginine,glutamine, or asparagine: or X₉ is plienylalanine, tryptophan, ortyrosine; or any combination of the foregoing.

In another specific embodiment, the peptide has the amino acid sequenceGX₅ X₆ HTX₇ CX₈ X₉ GX₁₀ SX₁₁ KPX₁₂ X₁₃ NCX₁₄ X₁₅ X₁₆ X₁₇ X₁₈ (SEQ IDNO:32), wherein X₅ is glycine or deleted, X₆ is any amino acid residue,X₇ is an amino acid residue with an aliphatic side chain, X₈ is a polaramino acid residue, X₉ is an amino acid residue with an aromatic sidechain, X₁₀ is a polar amino acid residue, X₁₁ is any amino acid residue,X₁₂ is a polar amino acid residue or deleted, X₁₃ is a basic amino acidresidue or deleted, X₁₄ is a small chain amino acid residue, X₁₅ is anyamino acid residue, X₁₆ is a basic or polar neutral amino acid residue,X₁₇ is any amino acid residue, and X₁₈ is an aliphatic amino acidresidue. More specifically, X₆ is isoleucine, leucine, threonine, orvaline; X₇ is alanine, valine, isoleucine, or leucine; X₈ is lysine,arginine, glutamine, or asparagine; X₉ is phenylalanine, tryptophan, ortyrosine; X₁₀ is threonine, serine, aspartic acid, or glutamic acid; X₁₁is leucine, isoleucine, methionine, or threonine; X₁₂ is serine ordeleted; X₁₃ is lysine, arginine, or deleted; X₁₄ is glycine or alanine;X₁₅ is asparagine, glutamine, glycine, serine, arginine, or lysine; X₁₆is lysine, arginine, asparagine, or glutamine; X₁₇ is lysine, arginine,isoleucine, leucine, or valine; or X₁₈ is alanine, valine, isoleucine,or leucine; or any combination of the foregoing.

The present invention also advantageously provides for combining twooverlapping peptides into a larger peptide having up to 35 amino acidresidue. In one such embodiment, the peptide has the amino acid sequenceX₁ BYCKIX₂ CX₃ X₄ GX₅ X₆ HTX₇ CX₈ X₉ GX₁₀ SX₁₁ KPX₁₂ X₁₃ NCX₁₄ X₁₅ X₁₆X₁₇ X₁₈ (SEQ ID NO:33), wherein X₁ is a neutral amino acid residue, X₂is a basic amino acid residue or deleted, X₃ is any amino acid residue,X₄ is a polar amino acid, X₅ is glycine or deleted, X₆ is any amino acidresidue, X₇ is an amino acid residue with an aliphatic side chain, X₈ isa polar amino acid residue, X₉ is an amino acid residue with an aromaticside chain, X₁₀ is a polar amino acid residue, X₁₁ is any amino acidresidue, X₁₂ is a polar amino acid residue or deleted, X₁₃ is a basicamino acid residue or deleted, X₁₄ is a small chain amino acid residue,X₁₅ is any amino acid residue, X₁₆ is a basic or polar neutral aminoacid residue, X₁₇ is any amino acid residue, and X₁₈ is an aliphaticamino acid residue. In a more specific embodiment, X₁ is valine orasparagine; X₂ is lysine, arginine, or deleted; X₃ is proline, arginine,serine, or leucine; X₄ is arginine, lysine, or serine; X₆ isoleucine,leucine, threonine, or valine; X₇ is alanine, valine, isoleucine, orleucine; X₈ is lysine, arginine, glutamine, or asparagine; X₉ isphenylalanine, tryptophan, or tyrosine; X₁₀ is threonine, serine,aspartic acid, or glutamic acid; X₁₁ is leucine, isoleucine, methionine,or threonine; X₁₂ is serine or deleted; X₁₃ is lysine, arginine, ordeleted; X₁₄ is glycine or alanine; X₁₅ is asparagine, glutamine,glycine, serine, arginine, or lysine; X₁₆ is lysine, arginine,asparagine, or glutamine; X₁₇ is lysine, arginine, isoleucine, leucine,or valine; or X₁₈ is alanine, valine, isoleucine, or leucine; or anycombination of the foregoing.

In another specific embodiment, the peptide has the amino acid sequenceKX₁₉ X₂₀ IX₂₁ X₂₂ X₂₃ HNX₂₄ FRQKX₂₅ AX₂₆ GLE (SEQ ID NO:34), wherein X₁₉is a basic or neutral polar amino acid residue X₂₀ is any amino acidresidue X₂₁ is an aliphatic amino acid residue, X₂₂ is a polar aminoacid residue, X₂₃ is a polar charged amino acid residue, X₂₄ is a polaramino acid residue, X₂₅ is an aliphatic amino acid residue; and X₂₆ is apolar basic or neutral amino acid residue. More specifically, X₁₉ isasparagine, glutamine, or lysine; X₂₀ is aspartic acid, glutamic acids,leucine, or isoleucine; X₂₁ is valine, leucine, or isoleucine; X₂₂arginine, asparagine, or serine; X₂₃ is glutamic acid or arginine; X₂₄is aspartic acid, glutamic acid, asparagine, glutamine, or arginine; X₂₅is valine, leucine, or isoleucine; or X₂₆ is arginine, asparagine, orlysine; or any combination of the foregoing.

In still another specific embodiment, the peptide has the amino acidsequence FRQKX₂₅ AX₂₆ GLETRGX₂₇ PGPQPX₂₈ (SEQ ID NO:35), wherein X₂₅ isan aliphatic amino acid residue, X₂₆ is a polar basic or neutral aminoacid residue, X₂₇ is a polar basic or neutral amino acid residue, andX₂₈ is any amino acid residue. More specifically, X₂₅ is valine,leucine, or isoleucine; X₂₆ is arginine, asparagine, or lysine; X₂₇ isasparagine or lysine; or X₂₈ is glycine, alanine, or proline; or anycombination of the foregoing.

As noted above, overlapping peptides may be provided as a single, largerpeptide. Thus, in another embodiment, the peptide has the amino acidsequence KX₁₉ X₂₀ IX₂₁ X₂₂ X₂₃ HNX₂₄ FRQKX₂₅ AX₁₆ GLETRGX₂₇ PGPQPX₂₈(SEQ ID NO:36), wherein X₁₉ is a basic or neutral polar amino acidresidue X₂₀ is any amino acid residue X₂₁ is an aliphatic amino acidresidue, X₂₂ is a polar amino acid residue, X₂₃ is a polar charged aminoacid residue, X₂₄ is a polar amino acid residue, X₂₅ is an aliphaticamino acid residue, X₂₆ is a polar basic or neutral amino acid residue,X₂₇ is a polar basic or neutral amino acid residue, and X₂₈ is any aminoacid residue. More specifically, X₁₉ is asparagine, glutamine, orlysine; X₂₀ is aspartic acid, glutamic acids, leucine, or isoleucine;X₂₁ is valine, leucine, or isoleucine; X₂₂ arginine, asparagine, orserine; X₂₃ is glutamic acid or arginine; X₂₄ is aspartic acid, glutamicacid, asparagine, glutamine, or arginine; X₂₅ is valine, leucine, orisoleucine; X₂₆ is arginine, asparagine, or lysine; X₂₇ is asparagine orlysine; or X₂₈ is glycine, alanine, or proline; or any combination ofthe foregoing.

In yet another embodiment, the peptide has the amino acid sequenceDELAX₂₉ X₃₀ AQX₃₁ WAX₃₂ QCX₃₃ X₃₄ X₃₅ X₃₆ HD (SEQ ID NO:37), wherein X₂₉is any amino acid residue, X₃₀ is an aliphatic amino acid residue, X₃₁is any amino acid residue, X₃₂ is a polar neutral amino acid residue,X₃₃ is a polar neutral or acidic amino acid residue, X₃₄ is an aromaticamino acid residue, X₃₅ is an aliphatic amino acid residue or deleted,and X₃₆ is any amino acid residue. More specifically, X₂₉ is tyrosine,lysine, or histidine; X₃₀ is isoleucine, leucine, or valine; X₃₁ isthreonine or valine; X₃₂ is serine or asparagine; X₃₃ is glutamine,asparagine, aspartic acid, or serine; X₃₄ is phenylalanine to tyrosine;X₃₅ is isoleucine, leucine, or deleted; or X₃₆ is glycine, asparagine,or valine; or any combination of the foregoing.

In still another specific embodiment, the peptide has the amino acidsequence NX₃₇ X₃₈ X₃₉ X₄₀ X₄₁ X₄₂ X₄₃ X₄₄ BX₄₅ FX₄₆ KX₄₇ GHYTQM (SEQ IDNO:38), wherein X₃₇ is a cyclic amino acid residue, X₃₈ is a polar basicor neutral amino acid residue, X₃₉ is any amino acid residue, X₄₀ is anyamino acid residue, X₄₁ is a non-polar amino acid residue, X₄₂ is anyamino acid residue, X₄₃ is a polar amino acid residue or glycine, X₄₄ isa polar basic or neutral amino acid residue, X₄₅ is a polar neutralamino acid or deleted, X₄₆ is any amino acid residue, and X₄₇ is anon-polar amino acid residue. More specifically, X₃₇ is proline ortyrosine; X₃₈ is asparagine, arginine, or histidine; X₃₉ is lysine,threonine, or valine; X₄₀ is aspartic acid, glycine, or lysine; X₄₁ isisoleucine, leucine, phenylalanine, or threonine; X₄₂ is glutamine,isoleucine, leucine, methionine, serine, or threonine; X₄₃ isasparagine, glutamic acid, histidine, lysine, or glycine, X₄₄ isaspartic acid, asparagine, glutamine, or serine; X₄₅ is asparagine ordeleted, X₄₆ is alanine, arginine, leucine, isoleucine, or serine; orX₄₇ is isoleucine, leucine, threonine, or valine; or any combination ofthe foregoing.

In still another embodiment, the peptide has the amino acid sequenceFX₄₆ KX₄₇ GHYTQMVWX₄₈ X₄₉ T (SEQ ID NO:39), wherein X₄₆ is any aminoacid residue. X₄₇ is a non-polar amino acid residue, X₄₈ is a small sidechain amino acid residue, and X₄₉ is a polar basic or neutral amino acidresidue. More specifically, X₄₆ is alanine, arginine, leucine,isoleucine, or serine; X₄₇ is isoleucine, leucine, threonine, or valine;X₄₈ is glycine or alanine; or X₄₉ is lysine or asparagine; or anycombination of the foregoing.

In another embodiment of the invention wherein two overlapping peptidesare combined, the peptide has the amino acid sequence NX₃₇ X₃₈ X₃₉ X₄₀X₄₁ X₄₂ X₄₃ X₄₄ BX₄₅ FX₄₆ KX₄₇ GHYTQMVWX₄₈ X₄₉ T (SEQ ID NO:40), whereinX₃₇ is a cyclic amino acid residue, X₃₈ is a polar basic or neutralamino acid residue, X₃₉ is any amino acid residue, X₄₀ is any amino acidresidue, X₄₁ is a non-polar amino acid residue, X₄₂ is any amino acidresidue, X₄₃ is a polar amino acid residue or glycine, X₄₄ is a polarbasic or neutral amino acid residue, X₄₅ is a polar neutral amino acidor deleted, X₄₆ is any amino acid residue, X₄₇ is a non-polar amino acidresidue, X₄₈ is a small side chain amino acid residue, and X₄₉ is apolar basic or neutral amino acid residue. More specifically, X₃₇ isproline or tyrosine, X₃₈ is asparagine, arginine, or histidine; X₃₉ islysine, threonine, or valine; X₄₀ is aspartic acid, glycine, or lysine;X₄₁ is isoleucine, leucine, phenylalanine, or threonine; X₄₂ isglutamine, isoleucine, leucine, methionine, serine, or threonine; X₄₃ isasparagine, glutamic acid, histidine, lysine, or glycine, X₄₄ isaspartic acid, asparagine, glutamine, or serine; X₄₅ is asparagine ordeleted, X₄₆ is alanine, arginine, leucine, isoleucine, or serine; X₄₇is isoleucine, leucine, threonine, or valine; X₄₈ is glycine or alanine;or X₄₉ is lysine or asparagine; or any combination of the foregoing.

In still another embodiment, the peptide has the amino acid sequence X₅₀ZX₅₁ X₅₂ X₅₃ X₅₄ X₅₅ HYLX₅₆ CNYGPX₅₇ GNX₅₈ X₅₉ X₆₀ (SEQ ID NO:41),wherein X₅₀ is a non-polar amino acid, X₅₁ is a polar acidic or neutralamino acid residue, X₅₂ is a polar basic or neutral amino acid residue,X₅₃ is a non-polar amino acid residue, X₅₄ is a moderately polar aminoacid residue, X₅₅ is a polar basic or neutral amino acid residue, X₅₆ isan aliphatic amino acid residue, X₅₇ is any amino acid residue, X₅₈ isany amino acid residue, X₅₉ is any amino acid residue, and X₆₀ is anyamino acid residue. More specifically, X₅₀ is isoleucine, leucine, ormethionine; X₅₁ is asparagine, aspartic acid, glutamine, or glutamicacid; X₅₂ is asparagine or lysine; X₅₃ is methionine or tryptophan; X₅₄glutamine, histidine, or tyrosine; X₅₅ is asparagine, lysine, orthreonine, X₅₆ isoleucine, leucine, or valine; X₅₇ is alanine, glycine,or serine; X₅₈ is aspartic acid, phenylalanine, or tyrosine; X₅₉ isglutaminie, leucine, isoleucine, methionine, or phenylalanine; or X₆₀ isasparagine, aspartic acid, or glycine; or any combination of theforegoing.

In specific embodiments, the invention is directed to a peptide havingan amino acid sequence selected from the group consistingof:NNYCKIKCRKGIHTLCKFGT (SEQ ID NO: 8), oritshomolog;GIHTLCKFGTSMKPNCGRNV (SEQ ID NO:9), oritshomolog;NNYCKIKCRKGIHTLCKFGTGTSMKPNCGRNV (SEQ ID NO:42),or itshomolog;KNEILKRHNDFRQNVAKGLE (SEQ ID NO:12), oritshomolog;FRQNVAKGLETRGKPGPQPP (SEQ ID NO:13), oritshomolog;KNEILKRHNDFRQNVAKGLETRGKPGPQPP (SEQ ID NO:43), oritshomolog;DELAKIAQTWANQCDFNHDD (SEQ ID NO:16), oritshomolog;NYKVGLQNSNFRKVGHYTQM (SEQ ID NO:22), oritshomolog;FRKVGHYTQMVWGKT (SEQ ID NO:23), or itshomolog;NYKVGLQNSNFRKVGHYTQMVWGKT (SEQ ID NO:44), or itshomolog;andIEDNWYTHYLVCNYGPGGND (SEQ ID NO:26), or itshomolog;

In specific embodiments, infra, peptides having SEQ ID NOS:8, 9, 12, 13,16, 22, 23, and 26 are shown to be dominant in more than one strain ofmouse.

In addition to homologous variants, allelic variant, consensus variant,and combined variant peptides of the invention, the present inventionfurther provides a recombinant polypeptide comprising two or morepeptides non-contiguously arranged relative to the native sequence ofvespid venom antigen 5. Although the term recombinant generally refersto polypeptides generated by genetic engineering, which meaning is fullyintended according to the present invention, the term recombinant isused herein more generally, to refer to polypeptides created bycombination (or recombination) of non-contiguous immunodominant peptidefragment of vespid venom antigen 5. Such fragments may be from antigen 5from the same species, antigen 5 peptides from different species,consensus antigen 5 peptides as described above, or any combination ofthe foregoing. In a specific embodiment, the peptides are selected fromthe group consisting of:NNYCKIKCRKGIHTLCKFGT (SEQ ID NO: 8), oritshomolog;GIHTLCKFGTSMKPNCGRNV (SEQ ID NO:9), oritshomolog;NNYCKIKCRKGIHTLCKFGTGTSMKPNCGRNV (SEQ ID NO:42),or itshomolog;KNEILKRHNDFRQNVAKGLE (SEQ ID NO:12), oritshomolog;FRQNVAKGLETRGKPGPQPP (SEQ ID NO:13), oritshomolog;KNEILKRHNDFRQNVAKGLETRGKPGPQPP (SEQ ID NO:43), oritshomolog;DELAKIAQTWANQCDFNHDD (SEQ ID NO:16), oritshomolog;NYKVGLQNSNFRKVGHYTQM (SEQ ID NO:22), oritshomolog;FRKVGHYTQMVWGKT (SEQ ID NO:23), or itshomolog;NYKVGLQNSNFRKVGHYTQMVWGKT (SEQ ID NO:44), or itshomolog;andIEDNWYTHYLVCNYGPGGND (SEQ ID NO:26), or itshomolog;

In specific embodiments, the present invention is directed to thefollowing homologous T cell peptides of hornet Ag5 from two species eachof hornet (Dol a and Dol m), yellowjacket (Ves m and Ves v) and wasp(Pol a and Pol e) and one species of fire ant (Sol i) shown in Table 2.As noted above, there are two forms of Dol m 5.01 and 5.02, referred toas Dol m A and B here. Residues given below refer to Dol m 5.02numbering, and gaps are added to obtain maximal sequence alignment.Where immunodominant peptides overlap, the fragment containing theoverlapping sequences are provided, with the specific sequencesdemarcated by a line under the peptide group. In such specificembodiments, the present invention contemplates both the individualimmunodominant peptides and the larger peptide containing theoverlapping segments.

                                      TABLE 2                                     __________________________________________________________________________    Homologs of Immunodominant Peptides                                           __________________________________________________________________________    Peptides 1 and 2 (residue 1-20 and 11-30)                                     Ves m                                                                             NNYCKI                                                                             KCLKGG                                                                             VHTACKYG  SLKP                                                                          NCGNKkV                                                                             SEQ ID NO: 45                                   Ves v                                                                             NNYCKI                                                                             KCLKGG                                                                             VHTACKYG  SLKP                                                                          NCGNKvV                                                                             SEQ ID NO: 46                                   Dol a                                                                             NNYCKI                                                                             CpKG tHTLCKYGTSMKP                                                                           NCGgKIVK                                                                            SEQ ID NO: 47                                   Dol mA                                                                            NNYCKI                                                                             KCsrG                                                                              IHTLCKFGTSMKP                                                                           NCGsKIVK                                                                            SEQ ID NO: 48                                   Dol mB                                                                            NNYCKI                                                                             KCrkG                                                                              IHTLCKFGTSMKP                                                                           NCGrnVVK                                                                            SEQ ID NO: 49                                   Pol a                                                                             VDYCKI                                                                             KCPSG                                                                              IHTVCQYGESTKPSKNCAGKVIK                                                                       SEQ ID NO: 50                                   Pol e                                                                             VDYCKI                                                                             KCPSG                                                                              IHTVCQYGESTKPSKNCAGKVIK                                                                       SEQ ID NO: 51                                   Sol i                                                                             YNYCNLQSCKRNNAIHTMCQY  TSPTPGPMCLEYSN                                                                   SEQ ID NO: 52                                   __________________________________________________________________________               2                                                                           2a                                                                   Peptides 5 and 6 (residue 41-60 and 51-70)                                    Ves m                                                                              KQDILKEHNDFRQKIARGLETRGNPGPQPPA                                                                       SEQ ID NO: 53                                    Ves v                                                                              KQDILKEHNDFRQKIARGLETRGNPGPQPPA                                                                       SEQ ID NO: 54                                    Dol a                                                                              KNEIVKRHNEFRQKVAqGLETRGNPGPQPPA                                                                       SEQ ID NO: 55                                    Dol mA                                                                             KNEIVnRHNQFRQKVAKGLETRGNPGPQPPA                                                                       SEQ ID NO: 56                                    Dol mB                                                                             KNEIlkRHNDFRQnVAKGLETRGkPGPQPPA                                                                       SEQ ID NO: 57                                    Pol a                                                                              KKLIVSEHNRFRQKVAQGLETRGNPGPQPAA                                                                       SEQ ID NO: 58                                    Pol e                                                                              KKLIVSEHNRFRQKVAQGLETRGNPGPQPAA                                                                       SEQ ID NO: 59                                    Sol i                                                                              KDAIVNKHNELRQRVASGKEMRGTNGPQPPA                                                                       SEQ ID NO: 60                                    __________________________________________________________________________                  6                                                               Peptide 9 (residue 81-100)                                                    Ves m DELAYiAQVWANQCQY                                                                             GHDT  SEQ ID NO:61                                       Ves v DELAYvAQVWANQCQY                                                                             GHDT  SEQ ID NO:62                                       Dol a DELAKIAQTWANQCnF                                                                             GHDQ  SEQ ID NO:63                                       Dol mA                                                                              DELAKIAQTWANQCsF                                                                             GHDQ  SEQ ID NO:64                                       Dol mB                                                                              DELAKIAQTWANQCdF                                                                             nHDD  SEQ ID NO:16                                       Pol a DELAHIAQVWASQCQFL                                                                            VHDK  SEQ ID NO:65                                       Pol e DELAHIAQVWASQCQFL                                                                            VHDK  SEQ ID NO:66                                       Sol i PELATIAQRWANQCTE                                                                             EHDA  SEQ ID NO:67                                       __________________________________________________________________________           9                                                                      Peptide 15 and 20 (residue 141-160 and 151-165)                               Ves m NPKKKFSeNn                                                                              FLKiGHYTQMVWANT                                                                            SEQ ID NO: 68                                    Ves v NPKKKFSgNd                                                                              FLKtGHYTQMVWANT                                                                            SEQ ID NO: 69                                    Dol a NPhKdlmhNN                                                                              FSKVGHYTQMVWGKT                                                                            SEQ ID NO: 70                                    Dol mA                                                                              NPKKGtigdnn                                                                             FSKVGHYTQMVWGKT                                                                            SEQ ID NO: 71                                    Dol mB                                                                              NYKvGlqnsN                                                                              FrKVGHYTQMVWGKT                                                                            SEQ ID NO: 44                                    Pol a NYNTGITKQN                                                                              FAKIGHYTQMVWGKT                                                                            SEQ ID NO: 72                                    Pol e NYNTGITKQN                                                                              FAKIGHYTQMVWGKT                                                                            SEQ ID NO: 73                                    Sol i NYNTGISFPSDDNILMKVEHYTQIVWAKT                                                                        SEQ ID NO: 74                                    __________________________________________________________________________    15                                                                                            20                                                            Peptide 18 (residue 176-195)                                                  Ves m   IQEnWHKHYLVCNYGPSGNF                                                                            SEQ ID NO: 75                                       Ves v   IQEKWHKHYLVCNYGPSGNF                                                                            SEQ ID NO: 76                                       Dol a   IENKWHTHYLVCNYGPAGNY                                                                            SEQ ID NO: 77                                       Dol mA  IENNWHTHYLVCNYGPAGNY                                                                            SEQ ID NO: 78                                       Dol mB  IEdNWyTHYLVCNYGPgGNd                                                                            SEQ ID NO: 26                                       Pol a   mENNMQNHYLICNYGPAGNY                                                                            SEQ ID NO: 79                                       Pol e   iENkMQNHYLICNYGPAGNY                                                                            SEQ ID NO: 80                                       Sol i   EPDNWTKHYLVCNYGPAGNV                                                                            SEQ ID NO: 81                                       __________________________________________________________________________

The present invention further provides a pharmaceutical composition fortreating vespid venom sensitivity, preferably where sensitivity toantigen 5 has been demonstrated, comprising any of the foregoingpeptides of the invention and a pharmaceutically acceptable carrier. Ina preferred aspect, the pharmaceutical composition comprises more thanone peptide of the invention, thus greatly increasing the breadth of itseffectiveness.

The invention naturally extends as well to a method for treatingsensitivity to vespid venom comprising administering to a vespid venomallergic patient a therapeutically effective amount of a pharmaceuticalcomposition of the invention. Preferably, the patient has beenidentified as sensitive to vespid venom antigen 5.

As demonstrated herein, the peptides of the invention are cross-reactivefor a native testes protein. Thus, an important advance of the presentinvention is that it provides peptides specific for treatment of antigen5-sensitive patients, regardless of the cause of this antigen 5sensitivity In other words, as demonstrated herein, whether a subjectdevelops antigen 5 sensitivity from vespid stings, fire ant bites, or totestes protein, the peptides of the present invention provide anadvantageous therapeutic agent for treating such sensitivity.

Thus, a primary object of the present invention is to provideimmunodominant peptides from vespid venom antigen 5.

A corollary object is to provide such immunodominant peptides thatproduce T cell anergy in subjects sensitive or allergic to vespid venomantigen 5.

Yet another object of the invention is to provide combinations of suchimmunodominant peptides for a broad treatment of vespid venom sensitivepatients from a wide variety of MHC (HLA) backgrounds.

Still another object of the invention is to provide consensusimmunodominant T cell epitopes of vespid venom antigen 5s.

These and other objects of the present invention can be betterappreciated by reference to the following drawings, the DetailedDescription of the Invention, and the Examples.

    ______________________________________                                        ABBREVIATIONS                                                                 ______________________________________                                        Ag5, Ag 5        antigen 5                                                    Dol m Dolichovespula maculata                                                                  white face hornet                                            Dol a D. arenaria                                                                              yellow hornet                                                Pol a Polistes annularis                                                                       wasp                                                         Pol e P. exclamans                                                                             wasp                                                         Ves m Vespula maculifrons                                                                      yellowjacket                                                 Ves v V. vulgaris                                                                              yellowjacket                                                 n-, r-           natural, recombinant (respectively)                          PCR              polymerase chain reaction                                    TCR              T cell receptor for antigen                                  tpx              testes protein                                               ______________________________________                                    

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Amino acid sequence of vespid antigen 5s. Antigen 5s wereisolated from yellowjackets (Vespula maculifrons and vulgaris), hornets(Dolichovespula areniaria and maculata) and wasps (Polistes annularisand exclamans). The sequences given are in the order of Ves m V, Ves vV, Dol a V, Dol m VA and VB, Pol a V, and Pol e V respectively. Residueswhich are common to all sequences are given on separate lines beneaththe sequences; residues underlined or dotted represent conserved orvariable regions, respectively.

FIG. 2. Peptide fragments of Dol m Antigen 5.2.

FIG. 3. Sequence similarity of hornet Ag5 and mouse and human tpx's.Bold characters indicate those residues of human and iuouse tpx's whichare identical with those of Ag5. The underlines regions of greater than7 residues in length have greater than 67% sequence identity, and eachis interspersed with fewer than 4 substituted residues.

FIG. 4. Stimulation index profile of BALB/c mice spleens after 4immunizations with natural (n)-Ag5. In vitro proliferation assays wereperformed with peptides at three concentrations: 1×10⁻⁵ M (open bar),1.25×10⁻⁶ M ("uphill" hatch), and 1.56×10⁻⁷ M ("downhill" hatch). Thecorresponding concentrations of recombinant antigen 5 control were1.0×10⁻⁶ M, 1.25×10⁻⁷, and 1.56×10⁻⁸ M. The blank was 2200 cpm.

FIG. 5. Stimulation index profile of BALB/c mice spleens after 4immunizations with recombinant (r)-Ag5. In vitro proliferation assayswere performed with peptides at three concentrations: 1×10⁻⁵ M (openbar), 1.25×10⁻⁶ M ("uphill" hatch), and 1.56×10⁻⁷ M ("downhill" hatch).The corresponding concentrations of recombinant Antigen 5 were 1.0×10⁻⁶M, 1.25×10⁻⁷, and 1.56×10⁻⁸ M. The blank was 4700 cpm.

FIG. 6. Stimulation index profile of BALB/c mice spleens after 4immunizations with r-Ag5. In vitro proliferation assays were performedwith peptides at three concentrations: 1×10⁻⁵ M (open bar), 1.25×10⁻⁶ M("uphill" hatch), and 1.56×10⁻⁷ M ("downhill" hatch). The correspondingconcentrations of recombinant Antigen 5 were 1.0×10⁻⁶ M, 1.25×10⁻⁷, and1.56×10⁻⁸ M. The blank was 3970 cpm.

FIG. 7. Stimulation index profile of BALB/c mice spleens after 4immunizations with r-frag IN (residue 1-114 of antigen 5). In vitroproliferation assays were performed with peptides at threeconcentrations: 1×10⁻⁵ M (open bar), 1.25×10⁻⁶ M ("uphill" hatch), and1.56×10⁻ M ("downhill" hatch). The corresponding concentrations ofrecombinant Antigen 5 were 1.0×10⁻⁶ M, 1.25×10⁻⁷, and 1.56×10⁻⁸ M. Theblank was 4690 cpm.

FIG. 8. Stimulation index profile of BALB/c mice spleens after 4immunizations with r-frag C (residue 151-204 of antigen 5). In vitroproliferation assays were performed with peptides at threeconcentrations: 1×10⁻⁵ M (open bar), 1.25×10⁻⁶ M ("uphill" hatch), and1.56×10⁻⁷ M ("downhill" hatch). The corresponding concentrations ofrecombinant Antigen 5 were 1.0×10⁻⁶ M, 1.25×10⁻⁷, and 1.56×10⁻⁸ M. Theblank was 6620 cpm.

FIG. 9. Stimulation index profile of BALB/c mice spleens after 5immunizations with r-frag IN. In vitro proliferation assays wereperformed with peptides at three concentrations: 1×10⁻⁵ M (open bar),1.25×10⁻⁶ M ("uphill" hatch), and 1.56×10⁻ M ("downhill" hatch). Thecorresponding concentrations of recombinant Antigen 5 were 1.0 ×10⁻⁶ M,1.25×10⁻⁷, and 1.56×10⁻⁸ M. The blank was 4410 cpm.

FIG. 10. Stimulation index profile of BALB/c mice spleens after 5immunizations with r-frag C. In vitro proliferation assays wereperformed with peptides at three concentrations: 1×10⁻⁵ M (open bar),1.25×10⁻⁶ M ("uphill" hatch), and 1.56×10⁻⁷ M ("downhill" hatch). Thecorresponding concentrations of recombinant Antigen 5 were 1.0×10⁻⁶ M,1.25×10⁻⁷ and 1.56×10⁻⁸ M. The blank was 7520 cpm.

FIG. 11. Stimulation index profile of ASW/sn mice spleens after 5immunizations with r-Ag5. In vitro proliferation assays were performedwith peptides at three concentrations: 1×10⁻⁵ M (open bar), 1.25×10⁻⁶ M("uphill" hatch), and 1.56×10⁻⁷ M ("downhill" hatch). The correspondingconcentrations of recombinant Antigen 5 were 1.0×10⁻⁶ M, 1.25×10⁻⁷, and1.56×10⁻⁸ M. The blank was 5640 cpm.

FIG. 12. Stimulation index profile of P/J mice spleens after 5immunizations with r-Ag5. In vitro proliferation assays were performedwith peptides at three concentrations: 1×10⁻⁵ M (open bar), 1.25×10⁻⁶ M("uphill" hatch), and 1.56×10⁻⁷ M ("downhill" hatch). The correspondingconcentrations of recombinant Antigen 5 were 1.0 ×10⁻⁶ M, 1.25×10⁻⁷, and1.56×10⁻⁸ M. The blank was 1730 cpm.

FIG. 13. Stimulation index profile of C3H/He mice spleens after 5immunizations with r-Ag5. In vitro proliferation assays were performedwith peptides at three concentrations: 1×10⁻⁵ M (open bar), 1.25×10⁻⁶ M("uphill" hatch), and 1.56×10⁻⁷ M ("downhill" hatch). The correspondingconcentrations of recombinant Antigen 5 were 1.0×10⁻⁶ M, 1.25×10⁻⁷, and1.56×10⁻⁸ M. The blank was 3130 cpm.

FIG. 14. Stimulation index profile of CS7B1/6 mice spleens after 5immunizations with r-Ag5. In vitro proliferation assays were performedwith peptides at three concentrations: 1×10⁻⁵ M (open bar), 1.25×10⁻⁶ M("uphill" hatch), and 1.56×10 ⁻⁷ M ("downhill" hatch). The correspondingconcentrations of recombinant Antigen 5 were 1.0×10⁻⁶ M, 1.25×10⁻⁷, and1.56×10⁻⁸ M. The blank was 1050 cpm.

FIG. 15. Stimulation index profile of BALB/c mice spleen cells after 4immunizations with Tpx-N. Stimulation was tested with peptide or proteinat three different concentrations: Open bars--1.0×10⁻⁵ M peptide or FragIN, 2.5×10⁻⁶ M Tp X-N; "uphill" hatch--1.25×10⁻⁶ M peptide or Frag IN.3.13×10⁻⁷ M, Tpx-N; "downhill" hatch--1.5×10⁻⁷ M peptide and Frag IN,3.91×10⁻⁸ M Tpx-N; and stippled --1.96×10⁻⁸ M Frag IN, 4.89×10⁻⁹ MTpx-N. The blank was 7270 cpm.

FIG. 16. Stimulation index profile of BALB/c mice spleen cells after 4immunizations with Tpx-C. Stimulation was tested with peptide or proteinat three different concentrations: Open bars--1×10⁻⁵ M peptide, 5×10⁻⁶ MFrag C or Tpx-C; "uphill" hatch--1.25×10⁻⁶ M peptide; 6.25×10⁻⁷ M Frag Cor Tpx-C; "downhill" hatch--1.56×10⁻⁷ M peptide, 7.8×10⁻⁸ M Frag C,Tpx-C: and stippled--9.75×10⁻⁹ M Frag-C or Tpx-C. The blank was 5110cpm.

DETAILED DESCRIPTION OF THE INVENTION

As noted above, the present invention provides the sequences ofimmunodominant peptides of vespid venom antigen 5, and correspondingpeptides from other antigens, such as fire ant Sol i 3.

The term "immunodominant peptide" is used herein to refer to a peptidethat contains a T cell epitope of the antigen, such that T cellsimmunized with the antigen will be stimulated when contacted with thepeptide. The term T cell epitope is used herein in conformity with thedefinition in immunology. In one embodiment, an immunodominant epitopeis an epitope that induces a greater than about 4-fold increase instimulation of immunized T cells in an in vitro stimulation assay,particularly a greater than 2-fold, preferably greater than 4-fold, andmore preferably greater than 6-fold increase in the stimulation index ina ³ H-thymidine incorporation assay. In another embodiment, animmunodominant peptide is a peptide that induces proliferation orstimulation of different MHC-restricted populations of immunized Tcells.

An immunodominant peptide of the invention may contain one epitope, orit may contain more than one epitope.

An immunomodulatory peptide" is a peptide that affects immune responsein vivo. Preferably, an immunomodulatory peptide of the inventioninduces T cell anergy when administered to a subject.

The invention also provides pharmaceutical compositions effective forthe treatment of a vespid venom allergen-specific allergic conditioncomprising a polypeptide of the invention, and methods for treating suchallergic conditions comprising administering a therapeutically effectivedose of the pharmaceutical compositions of the invention.

The polypeptides of the invention can also be useful for diagnosis ofvespid venom-specific allergic conditions.

As used herein. the term "vespid venom allergen" refers to a proteinfound in the venom of a vespid, to which susceptible people aresensitized on exposure to the sting of the insect. While most antigensare characterized by being reactive with specific IgG class antibodies,an allergen is characterized by also being reactive with IgE typeantibodies. The IgE type antibodies are responsible for mediating thesymptoms of an allergic condition, i.e., immediate-typehypersensitivity. According to the invention, the vespid venom allergenof interest is vespid venom antigen 5.

As herein, the term "vespid" is used according to the practice of thosein the field of allergy, and refers to insects belonging to theworldwide family of Vespidae, i.e., social wasps including hornets,yellowjackets, and paper wasps. In particular, vespids include thesubfamilies Vespinae and Polistinae. More particularly, the vespidsinclude the genera Vespa Linnaeus, Vespula Thomson, DolichovespulaRohwer, and Polistes Latreille. Species in the genus Vespula include butare not limited to V. germanica (Fab.), V. squamosa (Drury), V.maculifrons (Buysson), V. flavopilosa (Jacobson), V. vulgaris (L.), andV. pensylvannica (Saussure). Species in the genus Polistes include butare not limited to P. annularis (Linnaeus), P. exclamans (Viereck), P.metricus (Say), P. fuscatus (Fabricius), and P. apachus (Saussure).Species in the genus Dolicliovespula include but are not limited to D.maculata (L.) and D. arenaria (Fab.). Species in the genus Vespa includebut are not limited to V. crabro (L.) and V. orientalis (Linnaeus).

As used herein, the term "immunomodulatory" refers to an ability toincrease or decrease an antigen-specific immune response, either at theB cell or T cell level. Immunomodulatory activity can be detected e.g.,in T cell proliferation assays, by measurement of antibody production,lymphokine production or T cell responsiveness. In particular, inaddition to affects on T cell responses, the immunomodulatorypolypeptides of the invention may bind to immunoglobulin (i.e.,antibody) molecules on the surface of B cells, and affect B cellresponses as well.

For the sake of clarity, the present invention is described in detail insections relating to an immunomodulatory fragment of a vespid venomantigen 5, or derivatives and analogs of the vespid venom antigen 5,assays with the immunomodulatory vespid venom antigen 5, fragments, orderivatives or analogs thereof, and finally therapeutic and diagnosticuses of the vespid venom antigen 5 fragments, or derivatives or analogsthereof.

Immunodominant Peptides of Vespid Venom Ag5

An immunodominant peptide of the invention is a peptide comprising atleast one T cell epitope, with a minimum size of approximately 8 aminoacid residues (the minimum length generally regarded as requisite forantigen-specific recognition by a T cell). A peptide comprising at leastone T cell epitope is capable of eliciting a T cell response. such asstimulation or T cell anergy (tolerization).

The immunodominant peptides of the invention have been identified inmurine backgrounds. However, those peptides that are antigenic (comprisea T cell epitope) in more than one mouse strain are excellent candidatesfor immunodominant epitopes in antigen 5 sensitive humans. As pointedout above, on the basis of findings with Fel d 1 and Der p 1, it isreasonable to conclude that the T cell epitope data of a vespid venomantigen 5 found in mice, particularly in multiple strains, will beapplicable for humans.

Also contemplated by the term "immunodominant peptide" are peptideswhich join two or more discontinuous epitopes (or the specific peptidesdisclosed herein) in a single polypeptide. In one aspect, such"aggregate" polypeptides can be prepared chemically, e.g., bycrosslinking peptides, or using condensation techniques to form peptidebonds. Alternatively, synthetic DNA molecules encoding such aggregatepeptides can be prepared, inserted in a recombinant expression vector,and the aggregate peptide expressed by host cells transformed ortransfected with the expression vector and cultured under conditionsthat allow for expression of the aggregate polypeptide.

Immunodominant peptides of the invention can also be prepared as fusionproteins.

Amino acids used for peptide synthesis may be standard Boc (N.sup.α-amino protected N.sup.α -t-butyloxycarbonyl) amino acid resin with thestandard deprotecting, neutralization, coupling and wash protocols ofthe original solid phase procedure of Merrifield [1963, J. Am. Chem.Soc. 85:2149-2154], or the base-labile N.sup.α -amino protected9-fluorenylmethoxycarbonyl (Fmoc) amino acids first described by Carpinoand Han [1972, J. Org. Chem. 37:3403-3409]. Both Fmoc and Boc N.sup.α-amino protected amino acids can be obtained from Fluka, Bachem,Advanced Chemtech, Sigma, Cambridge Research Biochemical, Bachem, orPeninsula Labs or other chemical companies familiar to those whopractice this art. In addition, the method of the invention can be usedwith other N.sup.α -protecting groups that are familiar to those skilledin this art. Solid phase peptide synthesis may be accomplished bytechniques familiar to those in the art and provided, for example, inStewart and Young, 1984, Solid Phase Synthesis, Second Edition, PierceChemical Co., Rockford, Ill.; Fields and Noble, 1990, Int. J. Pept.Protein Res. 35:161-214, or using automated synthesizers, such as soldby ABS.

Alternatively, immunodominanit peptides of the invention can be preparedby recombinant DNA techniques. Thus, in accordance with the presentinvention there may be employed conventional molecular biology,microbiology, and recombinant DNA techniques within the skill of theart. Such techniques are explained fully in the literature [see, e.g.,Sambrook, Fritsch & Maniatis, Molecular Cloning: A Laboratory Manuial,Second Edition (1989) Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y. [herein "Sambrook et al., 1989"]; DNA Cloning: A PracticalApproach, Volumes I and II [D. N. Glover ed. 1985]; OligonucleotideSynthesis [M. J. Gait ed. 1984]; Nucleic Acid Hybridization [B. D.Harnes & S. J. Higgins eds. (1985)]; Transcription And Translation ]B.D. Hames & S. J. Higgins, eds. (1984)]; Animal Cell Culture [R. I.Freshney, ed. (1986)]. Immobilized Cells And Enzymes [IRL Press,(1986)], B. Perbal, A Practical Guide To Molecular Cloning (1984): F. M.Ausubel et al. (eds.), Current Protocols in Molecular Biology, JohnWiley & Sons, Inc. (1994)].

Peptides comprising at least two regions, each region comprising atleast one T cell epitope of a antigen 5 are also within the scope of theinvention. Each region of such peptides is derived from the same or fromdifferent antigen 5. Isolated peptides, each comprising at least two Tcell epitopes of antigen 5, are particularly desirable for increasedtherapeutic effectiveness. Peptides immunologically related by T cellcross-reactivity, which are capable of reacting with the same T cells asa peptide of the invention, are also contemplated.

As noted above, isolated peptides of the invention can be produced byrecombinant DNA techniques in a host cell transformed with a nucleicacid having a sequence encoding such peptide. The isolated peptides ofthe invention can also be produced by chemical synthesis. In certainlimited situations, isolated peptides can be produced by chemicalcleavage of the protein allergen. When a peptide is produced byrecombinant techniques, host cells transformed with a nucleic acidhaving a sequence encoding the peptide or the functional equivalent ofthe nucleic acid sequence are cultured in a medium suitable for thecells and peptides can be purified from cell culture medium, host cells,or both using techniques known in the art for purifying peptides andproteins including ion-exchange chromatography, gel filtrationchromatography, ultrafiltration, electrophoresis, or imrnunopurificationwith antibodies specific for the peptide, antigen 5 from which thepeptide is derived, or a portion thereof. Isolated peptides of theinvention are substantially free of cellular material or culture mediumwhen produced by recombinant DNA techniques, or substantially free ofchemical precursors or other chemicals when synthesized chemically.

Naturally, the present invention provides expression vectors and hostcells transformed to express the peptides. A nucleic acid sequencecoding for a peptide of the invention may be expressed in bacterialcells such as E. coli, insect cells (baculovirus), yeast, or mammaliancells such as Chinese hamster ovary cells (CHO). Suitable expressionvectors, promoters, enhancers, and other expression control elements maybe found in Sambrook et al. Molecular Cloning: A Laboratory Manual,second edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y. (1989). Other suitable expression vectors, promoters, enhancers,and other expression elements are known to those skilled in the art.Many yeast and bacterial vectors are commercially available or ondeposit at various depositories. Baculovirus and mammalian expressionsystems are also available.

For expression in E. coli, suitable expression vectors include, amongothers, pTRC [Amann et al., Gene, 69:301-315, (1988)], pGEX (AmradCorp., Melbourne, Australia); pMAL (N.E. Biolabs. Beverly, Mass.); pRIT5(Pharmacia, Piscataway, N.J.); pET-11d (Novagen, Madison, Wis.) [Jameelet al., J. Virol. 64:3963-3966 (1990)]: pQE12 (QIAGEN, Chatsworth,Calif.): and pSEM [Knapp et al., BioTechniques, 8:280-281 (1990)]. Theuse of pTRC, and pET-111d, for example, will lead to the expression ofunfused protein. The use of pMAL, pRIT5, pSEM, pQE12, and pGEX will leadto the expression of allergen fused to maltose E binding protein (pMAL),protein A (pRIT5), truncated β-galactosidase (PSEM), hexahistidine(pQE12), or glutathione S-transferase (pGEX). When a polypeptideallergen is expressed as a fusion protein, it is particularlyadvantageous to introduce an enzymatic cleavage site at the fusionjunction between the carrier protein and a polypeptide. Theimmnunodominant polypeptide (harboring the peptide of the invention) maythen be recovered from the fusion protein through enzymatic cleavage atthe enzymatic site and biochemical purification using conventionaltechniques for purification of proteins and peptides. Suitable enzymaticcleavage sites include those for blood clotting Factor Xa or thrombinfor which the appropriate enzymes and protocols for cleavage arecommercially available from, for example, Sigma Chemical Company, St.Louis, Mo. and N.E. Biolabs, Beverly, Mass. The different vectors alsohave different promoter regions allowing constitutive or inducibleexpression with, for example, IPTG induction (PRTC, Amann et al.,(1988), supra; pET-11d. Novagen, Madison, Wis.) or temperature induction(pRIT5, Pharmacia, Piscataway, N.J.).

Human T cell stimulating activity can be tested by culturing T cellsobtained form an individual sensitive to vespid venom (i.e., anindividual who has an IgE mediated immune response to antigen 5) with animmunodominant peptide of the invention and determining whetherproliferation of T cells occurs in response to the peptide as measured,e.g., by cellular uptake of tritiated thymidine. Stimulation indices forresponses by T cells to peptides can be calculated as the maximum CPM inresponse to a peptide divided by the control CPM. A T cell stimulationindex (S.I.) equal to or greater than two times the background level isconsidered "positive." Positive results are used to calculate the meanstimulation index for each peptide for the ;roup of peptides tested.Peptides of this invention comprise at least one T cell epitope and havea mean T cell stimulation index of greater than or equal to 2.0.Preferably the S.I. value is greater than four. A peptide having a Tcell stimulation index of greater than or equal to 2.0, and morepreferably greater than or equal to 4.0, is considered useful as atherapeutic agent. Preferred peptides have a mean T cell stimulationindex of at least 2.5, more preferably at least 3.5, even morepreferably at least 4.0, and most preferably at least 5.0.

In addition, preferred peptides have a positivity index (P.I.) of atleast about 100, more preferably at least 150, even more preferably atleast about 200 and most preferably at least about 250. The positivityindex for a peptide is determined by multiplying the mean T cellstimulation index by the percent of individuals, in a population ofindividuals sensitive to vespid venom (e.g., preferably at least 9individuals, more preferably at least 16 individuals or more, morepreferably at least 29 individuals or more, or even more preferably atleast 30 individuals or more), who have T cells that respond to thepeptide. Thus, the positivity index represents both the strength of a Tcell response to a peptide (S.I.) and the frequency of a T cell responseto a peptide in a population of individuals sensitive to vespid venom.

In order to determine precise T cell epitopes by, for example, finemapping techniques, a peptide having T cell stimulating activity andthus comprising at least one T cell epitope as determined by T cellbiology techniques is modified by addition or deletion of amino acidresidues at either the amino or carboxy terminus of the peptide andtested to determine a change in T cell reactivity to the modifiedpeptide. If two or more peptides which share an area of overlap in thenative protein sequence are found to have T cell stimulating activity,preferably human T cell stimulating activity, as determined by T cellbiology techniques, additional peptides can be produced comprising allor a portion of such peptides and these additional peptides can berested by a similar procedure. Following this technique, peptides areselected and produced recombinantly or synthetically. Peptides areselected based on various factors, including the strength of the T cellresponse to the peptide (e.g., stimulation index), the frequency of theT cell response to the peptide in a population of individuals sensitiveto vespid venom, and the potential cross-reactivity of the peptide withother antigen 5 homologs. The physical and chemical properties of theseselected peptides (e.g. solubility, stability) are examined to determinewhether the peptides are suitable for use in therapeutic compositions orwhether the peptides require modification as described herein. Theability of the selected peptides or selected modified peptides tostimulate human T cells (e.g., induce proliferation, lymphokinesecretion) is determined.

Additionally, preferred peptides of the invention do not bindimmunoglobulin E (IgE), or bind IgE to a substantially lesser extentthan the protein allergen from which the peptide is derived binds IgE.The major complications of standard immunotherapy are IgE-mediateresponse such as anaphylaxis. Immunoglobulin E is a mediator ofanaphylactic reactions which result from the binding and cross-linkingof antigen to IgE on mast cells or basophils and the release ofmediators (e.g., histamine, serotonin, eosinophil chemotactic factors).Thus, anaphylaxis in a substantial percentage of a population ofindividuals sensitive to vespid venom could be avoided by the use inimmunotherapy of a peptide of peptides which do not bind IgE in asubstantial percentage (e.g., at least about 75%) of a population ofindividuals sensitive to vespid venom allergen, or if the peptide bindsIgE, such binding does not result in the release of mediators from mastcells or basophile. The risk of anaphylaxis could be reduced by the usein immunotherapy of a peptide or peptides which have reduced IgEbinding. Thus, peptides which have minimal IgE stimulating activity, aredesirable for therapeutic effectiveness. Minimal IgE stimulatingactivity refers to IgE production that is less than the amount of IgEproduction and/or IL-4 production stimulated by the native proteinallergen.

A peptide of the invention. when administered to a vespidvenom-sensitive individual. is capable of modify,ing the allergicresponse of the individual to the allergen. Particularly, peptides ofthe invention comprising at least one T cell epitope of antigen 5 whenadministered to an individual sensitive to vespid venom are capable ofmodifying T cell response of the individual to the allergen. As usedherein, modification of the allergic response of an individual to vespidvenom antigen 5 can be defined as non-responsiveness or diminution insymptoms upon exposure to antigen 5, as determined by standard clinicalprocedures [see e.g., Varney et al., British Medical Journal,302:265-169 (1990)] including diminution in antigen 5 induced asthmaticsymptoms. As referred to herein, a diminution in symptoms includes anyreduction in allergic response of an individual to the allergen afterthe individual has completed a treatment regimen with a peptide orprotein of the invention. This diminution may be subjective (i.e., thepatient feels more comfortable in the presence of the allergen).Diminution in symptoms can be determined clinically as well, usingstandard skin tests as is known in the art.

As a result of the work described herein, peptides derived from antigen5 comprising at least one T cell epitope can be produced. T cellepitopes are believed to be involved in initiation and perpetuation ofthe immune responses to allergen(s), which are responsible for theclinical symptoms of vespid venom. These T cell epitopes are thought totrigger early events at the level of the T helper cell by binding to anappropriate HLA (MHC) molecule on the surface of an antigen presentingcell and stimulating the relevant T cell subpopulation. These eventslead to T cell proliferation, lymphokine secretion, local inflammatoryreactions, the recruitment of additional immune cells to the site, andactivation of the B cell cascade leading to production of antibodies.One isotype of these antibodies, IgE, is fundamentally important in thedevelopment of allergic symptoms and its production is influenced earlyin the cascade of events, at the level of the T helper cell, by thenature of the Iymphokines secreted. A T cell epitope is the basicelement or smallest unit of recognition by a T cell receptor where theepitope comprises amino acid residues essential to receptor recognition.Amino acid sequences which mimic those of T cell epitopes and whichmodify the allergic response to vespid venom antigen 5 are within thescope of this invention.

Exposure of vespid venom allergic patients to peptides of the presentinvention may tolerize or anergize appropriate T cell subpopulationssuch that they become unresponsive to vespid venom allergen(s), e.g.,antigen 5, and do not participate in mounting an immune response uponsuch exposure. In addition, administration of a peptide of the presentinvention may modify the lymphokine secretion profile as compared withexposure to the naturally-occurring vespid venom allergin or portionthereof (e.g., result in a decrease of IL-4 and/or an increase in IL-2).Furthermore, exposure to a peptide of the invention may influence T cellsubpopulations which normally participate in a response to vespid venomallergen(s) such that these T cells are drawn away from the site(s) ofnormal exposure to the allergen (e.g., nasal mucosa, skin, and lung)towards the site(s) of therapeutic administration of the peptide. Thisredistribution of T cell subpopulations may ameliorate or reduce theability of an individual's immune system to mount an immune response atthe site of normal exposure to the antigen 4, resulting in a diminutionin allergic symptoms.

Isolated peptides of the invention comprise at least one T cell epitopeof vespid venom antigen 5 and accordingly, the peptide comprises atleast approximately eight amino acid residues of the protein allergen.For purposes of therapeutic effectiveness, therapeutic compositions ofthe invention preferably comprise at least two T cell epitopes ofantigen 5. Accordingly, isolated peptides of the invention preferablycomprise at least two T cell epitopes and accordingly, the peptidecomprises at least approximately nine amino acid residues, andpreferably at least 15 amino acid residues. Additionally, therapeuticcompositions of the invention preferably comprise a sufficientpercentage of the T cell epitopes of the entire protein allergen suchthat a therapeutic regimen of administration to the composition to anindividual sensitive to vespid venom, results in T cells of theindividual being tolerized to the protein allergen. Syntheticallyproduced peptides of the invention comprising up to approximately 35amino acid residues in length, and most preferably up to approximately20 amino acid residues in length are particularly desirable as increasesin length beyond this point may result in difficulty in peptidesynthesis as well as retention of an undesirable property (e.g.,immunoglobulin binding or enzymatic activity) due to maintenance ofconformational similarity between the peptide and the protein allergenfrom which it is derived.

Another embodiment of the present invention provides peptides comprisingat least two regions, each region comprising at least one T cell epitopeof vespid venom antigen 5 and accordingly, each region comprises atleast approximately seven amino acid residues. These peptides comprisingat least two regions can comprise as many amino acid residues as desiredand preferably comprise at least about 14, even more preferably about25, and most preferably at most about 35 amino acid residues of antigen5. Each region of such peptide preferably comprises up to 20 amino acidresidues in length, more preferably up to 15 residues in length and mostpreferably up to 10 amino acid residues in length as increases in lengthof a region may result in difficulty in peptide synthesis as well asretention of an undesirable property (e.g., immunoglobulin binding orenzymatic activity) due to maintenance of conformational similaritybetween the peptide and the protein allergen from which it is derived.If desired, the amino acid sequences of the regions can be produced andjoined by a linker to increase sensitivity to processing byantigen-presenting cells. Such linker can be any non-epitope amino acidsequence or other appropriate linking or joining agent. To obtainpreferred peptides comprising at least two regions, each comprising atleast one T cell epitope, the regions are arranged in a configurationdifferent from a naturally-occurring configuration of the regions in theallergen or a combination of different antigen 5s. For example, theregions containing T cell epitope(s) can be arranged in a contiguousconfiguration and can preferably be derived from the same antigen or acombination of antigen 5 homologs from different species. Noncontiguousis defined as an arrangement of regions containing T cell epitope(s)which is different from that of an amino acid sequence present in theprotein allergen from which the regions are derived. Furthermore, thenoncontiguous regions containing T cell epitopes can be arranged in anonsequenitial order (e.g., in an arrangement different from the orderof the arrangement found in the native protein allergen from which theregion containing T cell epitope(s) are derived).

The present invention further provides for modification orderivatization of peptides in a library. Modifications of peptides arewell known to one of ordinary skill, and include phosphorylation,carboxymethylation, and acylation. Modifications may be effected bychemical or enzymatic means.

It is also possible to modify the structure of a peptide of theinvention for such purposes as increasing solubility, enhancingtherapeutic or preventive efficacy, or stability (e.g., shelf life exvivo, and resistance to proteolytic degradation in vivo). A modifiedpeptide can be produced in which the amino acid sequence has beenaltered, such as by amino acid substitution, deletion, or addition, tomodify immunogenicity and/or reduce allergenicity, or to which acomponent has been added for the same purpose.

For example, a peptide can be modified so that it maintains the abilityto induce T cell anergy and bind MHC proteins without the ability toinduce a strong proliferative response or possibly, any proliferativeresponse when administered in immunogenic form. In this instance,critical binding residues for the T cell receptor can be determinedusing known techniques (e.g., substitution of each residue anddetermination of the presence or absence of T cell reactivity). Thoseresidues shown to be essential to interact with the T cell receptor canbe modified by replacing the essential amino acid with another,preferably similar amino acid residue (a conservative substitution)whose presence is shown to enhance, diminish but not eliminate, or notaffect T cell reactivity. In addition, those amino acid residues whichare not essential for T cell receptor interaction can be modified bybeing replaced by another amino acid whose incorporation may enhance,diminish or not affect T cell reactivity, but does not eliminate bindingto relevant MHC.

Additionally, peptides of the invention can be modified by replacing anamino acid shown to be essential to interact with the MHC proteincomplex with another, preferably similar amino acid residue(conservative substitution) whose presence is shown to enhance, diminishbut not eliminate, or not effect T cell activity. In addition, aminoacid residues which are not essential for interaction with the MHCprotein complex but which still bind the MHC protein complex can bemodified by being replaced by another amino acid whose incorporation mayenhance, not effect, or diminish but not eliminate T cell reactivity.Preferred amino acid substitutions for non-essential amino acidsinclude, but are not limited to, substitutions with alanine, glutamicacid, or a methyl amino acid.

Another example of a modification of peptides is substitution ofcysteine residues preferably with serine, threonine, leucine or glutamicacid to minimize dimerization via disulfide linkages.

In order to enhance stability and/or reactivity, peptides can also bemodified to incorporate one or more polymorphism in the amino acidsequence of a protein allergen resulting from natural allelic variationor from homologous peptides. Additionally, D-amino acids, non-naturalamino acids, or non-amino acid analogs can be substituted or added toproduce a modified peptide within the scope of this invention.Furthermore, peptides of the present invention can be modified using thepolyethylene glycol (PEG) method of A. Sehon and co-workers (Wie et al.,supra) to produce a peptide conjugated with PEG. In addition, PEG can beadded during chemical synthesis of a peptide of the invention.Modifications of peptides or portions thereof can also includereduction/alkylation [Tarr in: Methods of Protein Microcharacrerization,J. E. Silver ed., Humana Press: Clifton, N.J. pp. 155-194) (1986)];acylation [Tarr. supra], esterification [Tarr, supra], chemical couplingto an appropriate carrier [Mishell and Shiigi, eds., Selected Methods inCellular Immunology, WH Freeman: San Francisco, Calif. (1980); U.S. Pat.No. 4,939,239]; or mild formalin treatment [Marsh, Int. Arch. AllergyApp. Immunol., 42:199-215 (1971)].

To facilitate purification and potentially increase solubility ofpeptides of the invention, it is possible to add reporter group(s) tothe peptide backbone. For example, polyhistidine can be added to apeptide to purifv the peptide on immobilized metal ion affinitychromatography [Hochuli, E. et al., Bio/Technology 6:1321-1325 (1988)].In addition, specific endoprotease cleavage sites can be introduced, ifdesired, between a reporter group and amino acid sequences of a peptideto facilitate isolation of peptides free of irrelevant sequences. Inorder to successfully desensitize an individual to a protein antigen, itmay be necessary to increase the solubility of a peptide by addingfunctional groups to the peptide or by not including hydrophobic T cellepitopes or regions containing hydrophobic epitopes in the peptides.

To potentially aid proper antigen processing of T cell epitopes within apeptide, canonical protease sensitive sites can be recombinantly orsynthetically engineered between regions, each comprising at least one Tcell epitope. For example, charged amino acid pairs, such as KK or RR,can be introduced between regions within a peptide during recombinantconstruction of the peptide. The resulting peptide can be renderedsensitive to cathepsin and/or other trypsin-like enzymes cleavage togenerate portions of the peptide containing one or more T cell epitopes.In addition, such charged amino acid residues can result in an increasein solubility of a peptide.

Fatty acyl peptide derivatives may also be prepared. For example, andnot by way of limitation, a free amino group (N-terminal or lysyl) maybe acylated, e.g., myristoylated. In another embodiment an amino acidcomprising an aliphatic side chain of the structure --(CH₂)_(n) CH₃ maybe incorporated in peptides of the library. This and other peptide-fattyacid conjugates suitable for use in the present invention are disclosedin U.K. Patent GB-8809162.4, International Patent ApplicationPCT/AU89/00166.

Activity Assays With Peptides of the Invention

Numerous assays are known in immunology for evaluating theimmunomodulatory activity of an antigen. For example, the immunodominantvespid venom can be used in diagnostic assays for allergic diseases,which are described in detail, infra. In general, such peptides can betested for the ability to induce proliferation of T cells from allergicsubjects, without reacting with antibodies specific for antigen 5.Preferably, such antibodies that are not reactive with the peptides ofthe invention in the diagnostic assay are of the IgE class. It isimportant to note that natural allergen-specific antibodies have beenfound to bind weakly to denatured vespid venom allergens.

The peptides of the invention can be tested in a proliferation assay forT cell responses. Generally, lymphocytes from a sensitized host areobtained. The host can be a mouse that has been immunized with a vespidvenom antigen 5, or with a crossreactive protein, such as testesspecific protein. Using techniques that are well known in the art, Tlymphocyte response to the peptide can be measured in vitro. In aspecific embodiment, infra, T cell responses are detected by measuringincorporation of ³ H-thymidine, which increases with DNA synthesisassociated with proliferation. Cell proliferation can also be detectedusing an MTT assay [Mossman, J. Immunol. Methods, 65:55-63,(1983); Niksand Otto, J. Immunol. Methods, 130:140-151 (1990)]. Alternatively,lymphokine production assays can be practiced according to the presentinvention. In one embodiment, lymphokine production can be assayed usingimmunological or co-stimulation assays [see, e.g., Fehlner et al., J.Immunol., 146:799 (1991)] or using the ELISPOT technique [Czerkinsky, etal., J. Immunol. Methods, 110:29 (1988)]. Alternatively, mRNA forlymphokines can be detected, e.g., by amplification [see Brenner, etal., Biotechniques, 7:1096 (1989)] or in situ hybridization [see, e.g.,Kasaian and Biron, J. Immunol., 142:1287 (1989)]. Of particular interestare those individuals whose T cells produce lymphokines associated withIgE isotype switch events, e.g., IL-4 and IL-5 [Purkeson and Isakson, J.Exp. Med., 175:973-982 (1992)]. Also of interest are the peptidefragments of the vespid venom antigen 5 that contain epitopes recognizedby T cells involved in IgE switch events. Any method for detecting Tcell proliferation known in the art can be used with the immunodominantvespid antigen 5 peptides obtained according to the present invention.

Thus, in a preferred aspect, the peptides of the present invention canbe used in in vitro assays with peripheral blood lymphocytes or celllines derived from peripheral blood lymphocytes, obtained from vespidvenom antigen 5 sensitive individuals to detect secretion of lymphokinesordinarily associated with allergic responses, e.g., IL-4. Such assaysmay indicate which epitopes are responsible or associated with theallergic condition. In this way, specific epitopes responsible for Tcell responses associated with allergic response can be identified. Thesequences of such epitopes can be compared to other vespid venom antigen5s and to environmental or autologous proteins to determine if there aresequence similarities that suggest possible cross-reactivity. Thepeptides can be tested for the ability to induce T cell anergy, e.g., bymega-dose administration, modification to produce an epitope antagonist,administration in the absence of the appropriate costimulatory signals,and other methods thought to result in T cell anergy. Peptidescontaining such epitopes are ideal candidates for therapeutics.

In a further embodiment, the polypeptides of the invention can be useddirectly in assays to detect the extent of cross-reactivity with otherenvironmental proteins and/or homologous proteins, with which they sharesequence similarity. In particular, the immunodominant fragments of thevespid venom antigen 5s that have sequence similarity with suchenvironmental, and more particularly, homologous proteins can beevaluated for cross reactivity with antibodies or T cell specific forsuch proteins. In a specific embodiment, the cross reactivity of vespidvenom antigen 5s with human and mouse testes specific protein can beevaluated.

Diagnostic and Therapeutic Uses of the Peptides

The present invention identifies therapeutically relevant polypeptidefragment of vespid venom antigen 5. The invention contemplates use ofthe imnunoactive fragments of vespid venom antigen 5 for the preparationof diagnostic or therapeutic compositions, for the use in the diagnosisand therapy of vespid venom allergen-specific allergic conditions. Inparticular, vespid antigen 5 from Dolichovespula maculata (white-facehornet) (Dol m V), Dolichovespula arenaria (yellow hornet) (Dol a V),Vespula vulgaris (yellowjacket) (Ves v V), Vespula maculifrons(yellowjacket) (Ves m V), Polistes annularis (wasp) (Pol a V), andPolistes exclamans (wasp) (Pol e V) are contemplated for use indiagnosis and therapy according to the present invention. Other vespidspecies known to harbor antigen 5, and thus represent additional antigen5 species of the invention, include but are not limited to Vespa crabo(European hornet), V. flavopilosa (yellow jacket), V. germanica(yellowjacket), V. pennsylvannica (yellowjacket), V. squamosa(yellowjacket), V. vidue (yellowjacket), and P. fuscatus (paperwasp).

Diagnostic Methods

The present invention contemplates in vitro diagnostic assays onperipheral blood lymphocytes, as described supra. Such diagnostic assayscan give detailed information about the antigen 5-specific T cellresponses, the phenotype of the T cell response, and preferably the Tcell epitope of the antigen 5 involved in T cell responses. Theinimunodominant epitope and the epitope involved in IgE isotype classswitch events can be detected, if they are not the same. In particular,the T cell epitopes of vespid venom antigen 5s that stimulateproliferation and/or lymphokine secretion of T cells of a phenotypeassociated with IgE isotype class switching events can be identified fora specific individual, or for a class of individuals who share MHChaplotype or a predominant T cell receptor variable region expression,or both.

In vivo assays for allergenicity generally consist of skin pricksensitivity assays, in which serially diluted amounts of an allergen areadministered either subcutaneously or intradermally into a patient'sskin, and wheel and erythema reactions are detected. As with in vitroassays, the availability of pure venom antigen 5 peptides greatlyincreases the value of the results of the in vivo diagnostic assayssince cross-reactivity with impurities in extracts prepared from vespidvenom sacs, and uncertainty about the epitope specificity can beavoided.

Therapeutic Methods

Therapeutic compositions of the invention (see, infra) can be used inimmunotherapy, also referred to as hyposensitization therapy.Immunotherapy has proven effective in allergic diseases, particularinsect allergy. Allergens are administered parenterally over a longperiod of time in gradually increasing doses. Such therapy may beparticularly effective when the allergen or allergens to which thepatient is sensitive have been specifically identified and the therapyis targeted to those allergen(s). Thus, the availability of specific,pure vespid venom antigen 5 peptide(s) in large quantities is importantfor immunotherapy of allergy.

The present invention contemplates use of peptides containing at leastan immunomodulatory T cell epitope of a vespid venom antigen 5 to inducespecific T cell anergy to the vespid venom antigen 5. Suchimmunomodulatory peptide contains one or more immunodominant epitope ofantigen 5. A peptide comprising such a T cell epitope and lacking a Bcell epitope can be administered to a patient. As discussed supra, thepresence of B cell epitopes on an allergen can cause an undesirablesystemic reaction when the allergen is used for immunotherapy. Thus, aparticular advantage of the invention is the capability to provideallergen polypeptides that do not cause undesirable systemic effects.

In one embodiment, one or more peptides can be injected subcutaneouslyto decrease the T cell response to the entire molecule, e.g., asdescribed by Briner et al. [Proc. Natl. Acad. Sci. U.S.A., 90:7608-12(1993)]. In another embodiment, one or more peptide can be administeredintranasally to suppress allergen-specific responses in naive andsensitized subjects [see e.g., Hoyne et al., J. Exp. Med., 178:1783-88(1993)].

Administration of a vespid venom antigen peptide of the invention isexpected to induce anergy, resulting in cessation of allergen-specificantibody production or allergen-specific T cell response, or both, andthus, have a therapeutic effect.

In a preferred aspect of the invention, peptide-based therapy to induceT cell anergy is customized for each individual or a group ofindividuals. Using the diagnostic methods of the present invention, thespecific T cell epitope or epitopes of a vespid venom antigen 5 involvedin the allergic response can be identified. Peptides comprising theseepitopes can then be used in an individualized immunotherapy regimen.

For example, by analogy and based on the data shown in Table 4, infra, atreatment protocol for BALB/c mice could include peptides 1, 5, 6, 11,15, 20, and 18, while treatment of ASW/Sn mice might include peptides 6,7, and 18: treatment of C3H/He might include peptides 2, 5, and 15, andtreatment of P/J mice might include peptides 1, 9, and 15.

Administration of the therapeutic compositions of the present inventionto desensitize an individual can be carried out using known techniques.For example, one or more immunodominant peptides of the inventioncomprising at least one T cell epitope can be administered incombination with an appropriate diluent, a carrier, and/or an adjuvant.To induce T cell anergy in an individual, the therapeutic composition ispreferably administered in non-immunogenic form, e.g., it does notcontain adjuvant. Pharmaceutically acceptable diluents include salineand aqueous buffer solutions. Pharmaceutically acceptable carriersinclude polyethylene glycol [Wie et al., International Archives ofAllergy and Applied Immunology, 64:84-99 (1981)] and liposomes [Strejanet al., Journal of Neuroimmunology, 7:27 (1984)]. Such compositions willgenerally be administered by injection (subcutaneous, intravenous,etc.), oral administration (e.g., as in the form of a capsule),transdermal application, or transmucosal administration (e.g. nasal,buccal, or rectal administration). The therapeutic compositions of theinvention are administered to individuals sensitive to vespid venom atdosages and for lengths of time effective to reduce sensitivity (i.e.,reduce the allergic response) of the individual to vespid stings. Atherapeutically effective amount of one or more of the same or differenttherapeutic compositions can be administered simultaneously orsequentially to an individual sensitive to vespid venom. Effectiveamounts of the therapeutic compositions will vary according to factorssuch as the degree of sensitivity of the individual to vespid venom, theage, sex, and weight of the individual, and the ability of the peptideto stimulate a T cell response in the individual.

In yet another aspect of the present invention, a composition isprovided comprising at least two immunodominant peptides (e.g., aphysical mixture of at least two peptides), each comprising at least oneT cell epitope of antigen 5. The peptides are derived from the same orfrom different homologs of antigen 5. Such compositions can beadministered in the form a therapeutic composition with apharmaceutically acceptable carrier of diluent. A therapeuticallyeffective amount of one or more of such compositions can be administeredsimultaneously or sequentially to an individual sensitive to vespidvenom.

Transmucosal administration. According to the invention, anytransmucosal route of administration, including but not limited torectal, oral, vaginal, buccal, etc. can be employed. In particular, thepresent invention is directed to the following transmucosal routes ofadministration. It can be readily appreciated that any of thetransmucosal routes of administration may be enhanced by use of amucosal penetration enhancer. The term "mucosal penetration enhancer"refers to a reagent that increases the rate or facility of transmucosalpenetration of peptide, such as but not limited to, a bile salt, fattyacid, surfactant or alcohol. In specific embodiments, the permeationenhancer can be sodium cholate, sodium dodecyl sulphate, sodiumdeoxycholate, taurodeoxycholate, sodium glycocholate, dimethylsulfoxideor ethanol. Suitable penetration enhancers also include glycyrrhetinicacid (U.S. Pat. No. 5,112,804 to Kowarski) and polysorbate-80, thelatter preferably in combination with an non-ionic surfactant such asnonoxynol-9, laureth-9, poloxamer-124, octoxynol-9, or lauramide-DEA(European Patent EP 0 242 643 B1 by Stoltz). The selection of aparticular mucosal penetration enhancer may depend on thecharacteristics of the specific mucosa. These factors are addressed ingreater detail below.

Administration Via Suppositories. In another aspect, peptide isformulated in a matrix suitable for rectal (or vaginal) insertion, i.e.,in a suppository. The invention is not limited to any particularsuppository formulation. Indeed, many suppository formulations are knownin the art, e.g., as described in Remington's Pharmaceutical Sciences,Physician's Desk Reference, and U.S. Pharmacopeia.

Administration Via a Buccal Patch. According to the invention, peptidecan be formulated in a buccal patch for administration via the interiorof the cheek. It may be appreciated that a buccal patch constitutesanother form of transmucosal administration. The technology forpreparing buccal patch formulations is known in the art, e.g.,Remington's Pharmaceutical Sciences, supra.

Oral-Pharyngeal Administration. In yet another embodiment, peptide canbe formulated for oral-pharyngeal, including sublingual and transbuccal,administration. For example, peptide can be incorporated in a "candy"matrix, such as that described in U.S. Pat. No. 4,671,953, in a gumbase, or a lozenge. In another embodiment, the peptide can be formulatedin a capsule or pill form for sublingual placement. It is particularlycontemplated that peptide for oral-pharyngeal administration may beformulated with a flavor masking agent or coating. Many flavor maskingagents for use with oral pharmaceuticals are known in the art, and canbe selected for use with the present invention.

Oral Administration. In still a further embodiment, peptide can beformulated for oral administration via the stomach and intestinalmucosa. For oral administration, peptide can be administered in acarrier designed for drug release in either the stomach (an acidicenvironment), or the intestines, or both. Many capsules, pills, andmatrices for oral administration of a drug are known in the art, and canbe selected on the basis of compatibility with peptide, and the desiredpoint and rate of drug release by the ordinary skilled physician. Oraladministration of peptide may require higher dosages than other routesof administration to overcome the effects of first pass metabolism bythe liver.

Transdermal Administration. In a further embodiment, as noted above, thepresent invention is directed to cransdermal administration of peptide.Various and numerous methods are known in the art for transdermaladministration of a compound, e.g., via a transdermal patch. Thesemethods and associated devices provide for control of the rate andquantity of administration of a drug, and some allow for continuousmodulation of drug delivery. Transdermal patches are described in, forexample, U.S. Pat. No. 5,407,713, issued Apr. 18, 1995 to Rolando etal.; U.S. Pat. No. 5,352,456, issued Oct. 4, 1004 to Fallon et al.; U.S.Pat. No. 5,332,213 issued Aug. 9, 1994 to D'Angelo et al.; U.S. Pat. No.5,336,168, issued Aug. 9, 1994 to Sibalis; U.S. Pat. No. 5,290,561,issued Mar. 1, 1994 to Farhadieh et al.; U.S. Pat. No. 5,254,346, issuedOct. 19, 1993 to Tucker et al.; U.S. Pat. No. 5,164,189, issued Nov. 17,1992 to Berger et al.; U.S. Pat. No. 5,163,899, issued Nov. 17, 1992 toSibalis; U.S. Pat. Nos. 5,088,977 and 5,087,240, both issued Feb. 18,1992 to Sibalis; U.S. Pat. No. 5,008,110, issued Apr. 16, 1991 toBenecke et al.; and U.S. Pat. No. 4,921,475, issued May 1, 1990 toSibalis, the disclosure of each of which is incorporated herein bvreference in its entirety.

It can be readily appreciated that a transdermal route of administrationmay be enhanced by use of a dermal penetration enhancer, e.g., such asenhancers described in U.S. Pat. No. 5,164,189 (supra), U.S. Pat. No.5,008,110 (supra), and U.S. Pat. No. 4,879,119, issued Nov. 7, 1989 toAruga et al., the disclosure of each of which is incorporated herein byreference in its entirety.

Pharmaceutically Acceptable Compositions

The in vivo diagnostic or therapeutic compositions of the invention mayalso contain appropriate pharmaceutically acceptable carriers,excipients, diluents and adjuvants. As used herein, the term"pharmaceutically acceptable" preferably means approved by a regulatoryagency of a government, in particular the Federal government or a stategovernment, or listed in the U.S. Pharmacopeia or another generallyrecognized pharmacopeia for use in animals, and more particularly inhumans, although a pharmaceutically acceptable carrier may share theattributes of such approved or recognized carriers without having itselfbeen approved or recognized. Examples of suitable pharmaceuticalcarriers are provided in "Remington's Pharmaceutical Sciences" by E. W.Martin.

Such pharmaceutically acceptable carriers can be sterile liquids, suchas water and oils, including those of petroleum, animal, vegetable orsynthetic origin, such as peanut oil, soybean oil, mineral oil, sesameoil and the like. Water is a preferred carrier when the pharmaceuticalcomposition is administered intravenously. Saline solutions and aqueousdextrose and glycerol solutions can also be employed as liquid carriers,particularly for injectable solutions. Suitable pharmaceuticalexcipients include mannitol, human serum albumin (HSA), starch, glucose,lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel,magnesium carbonate, magnesium stearate, sodium stearate, glycerolmonostearate, talc, sodium chloride, dried skim milk, glycerol,propylene, glycol, water, ethanol and the like. These compositions cantake the form of solutions, suspensions, tablets, pills, capsules,powders, sustained-release formulations and the like.

Such compositions will contain an effective diagnostic or therapeuticamount of the active compound together with a suitable amount of carrierso as to provide the form for proper administration to the patient.While intravenous injection is a very effective form of administration,other modes can be employed, such as by injection, or by oral, nasal, orparenteral administration.

The invention will be further clarified by the following examples, whichare intended to be purely exemplary of the invention.

EXAMPLE 1 Mapping T Cell Epitopes of Dol M Ag5

In the present Example, a series of 15 to 20-residue peptides with10-residue overlaps to encompass the entire white faced hornet Ag5molecule (form 2) was synthesized. The sequences of these peptides aregiven in FIG. 2. These peptides were used for mapping T cell epitopes ofhornet Ag5 by their stimulation of specific murine spleen cellsimmunized with native or recombinant Antigen 5. These peptides were alsoused to study the cross reacting T cell epitopes of hornet antigen 5 anda mouse testis protein Tpx [Mizuku et al., Mammalian Genome, 3:274-280(1992)]. This cross reactivity is suggested by the partial sequenceidentity of hornet antigen 5 with mouse and human testis proteins (FIG.3).

Results

Antibody response in mice of different haplotypes to recombinant hornetAg5. Five strains of mice were immunized with recombinant (r) whitefaced hornet Ag5 and alum as adjuvant. Four strains were found to behigh antibody responders for r-white faced hornet Ag5: BALB/v byj,ASW/Sn, C3H/He, and P/J. Antibody responses were evaluated by directELISA [King et al., J. Immunol., 154:577 (1995)]. The antibody titers(the reciprocal of the dilution yielding 50% of maximum signalextrapolated from ELISA curves in wells coated with r-Dol m 5.02 orn-Dol m 5.01) of these sera for n-(natural) and r-Ag5 are given in Table3. One strain, C57B1/6, was found to be a poor antibody responder.

                  TABLE 3                                                         ______________________________________                                        Antibody for Mice of Different Strains                                        Immunized with Recombinant White faced hornet Ag5                             Mice                  Half maximal Ab titer*                                  Strain    Haplotype   r-Dol m 5.02                                                                            n-Dol m 5.01                                  ______________________________________                                        C57B1/6   b           1 × 10.sup.2                                                                      2 × 10.sup.1                            BALB/c byj                                                                              d           8 × 10.sup.4                                                                      4 × 10.sup.4                            C3H/He    k           6 × 10.sup.3                                                                      2 × 10.sup.3                            P/J       p           2 × 10.sup.4                                                                      1 × 10.sup.4                            ASW/Sn    s           1 × 10.sup.5                                                                      1 × 10.sup.5                            ______________________________________                                         *Sera are from week five bleeding after three intraperitoneal                 immunizations with 0.2 ml of 10 μg/ml rDol m 5.02 and alum (5 μg/ml     in 0.05 m sodium phosphate buffer, pH 6.2, on weeks 0, 2, and 4.         

T cell response in BALB/c mice to natural hornet Ag5 and its recombinantprotein or fragments. FIGS. 4 and 5 show the proliferative responses ofspleen cells from BALB/c mice immunized 4 times with n- and r-Ag5respectively. Nearly identical results show that r-Ag5 and n-Ag5 sharecommon continuous T cell epitopes. Previous studies have shown thatr-Ag5 lacks the discontinuous B cell epitopes of n-Ag5, as r-Ag5 is notproperly folded with the disulfide bonds of n-Ag5 [King et al., J.Immunol., 154:577-584 (1995)]. FIG. 6 shows the response of spleen cellsfrom BALB/c mice immunized 5 times with r-Ag5. Higher proliferativeresponses were observed than those in FIG. 5, where mice were immunized4 times with r-Ag5. The results in FIG. 4-6 together indicate thatpeptides 1, 5, 6, 11, 15, 20, and 18 contain the T cell epitopes of Ag5,if we assume that peptides have stimulation indices of ≧4 representpositive responses with a high degree of certainty.

FIGS. 7 and 8 show the proliferation data with spleen cells from miceimmunized 4 times with recombinant N-terminal and C-terminal fragmentsof antigen 5, labelled IN (residue 1-114) and C (residue 151-204),respectively. Similar but higher proliferation results were obtainedwith mice after 5 times immunization, as given in FIGS. 9 and 10. As tobe expected, spleen cells from r-fragment IN or C immunized mice werestimulated by peptides whose sequences are represented by the immunogen.There is one exception shown in FIG. 9: spleen cells from r-fragment IN(residue 1-114) immunized mice were stimulated by peptide 18 (residue176-195).

T cell response in mice of different haplotypes to recombinant hornetAg5. FIGS. 11 to 13 show the proliferative response of spleen cells formice of ASW/Sn, P/J, and C3H/He, respectively. The results for ASW/Snmice in FIG. 11 indicate that there are four peptides with stimulationindices of ≧4: peptides 6, 7, 9, and 18. The results for P/J mice inFIG. 12 show multiple peptides with stimulation indices of ≧4: peptides1, 2, 5, 9, 12, 13, 14, 15, 20, and 17. The data for C3H/He mice in FIG.12 indicate three peptides with stimulation indices of ≧4: peptides 2, 5and 15.

FIG. 14 shows the proliferation responses of spleen cells for C57B1/6mice. Several peptides had stimulation indices of ≧2, but only onepeptide, number 9, showed a stimulation index approaching 4. A lowerproliferation response compared to the other mouse strains was alsoobserved with r-Ag5 as a control. These data indicate that C57B/6 miceshowed poor T cell responses to white faced hornet Ag5. as contrasted tothe other 4 strains of mice tested (Table 4).

                  TABLE 4                                                         ______________________________________                                        Summary of proliferation indices of r-Ag5 specific spleen                     cells from mice of different strains on stimulation with the                  dominant T cell epitope peptides.sup.1                                        Stimulating.sup.2                                                                     Stimulation Index.sup.3                                               antigen BALB/c   ASW/Sn   C3H/He P/J    C57B1/6                               ______________________________________                                        Peptide                                                                             1     8.6      1.9    2.4    6.9    2.5                                       2     2.9      2.2    4.3    4.8    1.9                                       5     9.3      1.4    5.8    4.0    1.7                                       6     5.8      8.8    2.0    2.2    1.9                                       7     (0.9)    7.7    2.2    3.3    2.1                                       9     3.8      4.0    2.7    9.9    3.9                                       11    5.8      0.9    3.0    1.4    2.7                                       12    2.5      1.4    3.3    4.0    2.5                                       13    (1.3)    1.2    3.6    4.5    2.5                                       14    2.3      2.3    2.9    4.2    2.0                                       15    5.6      1.4    4.7    7.8    2.8                                       20    6.5      1.7    2.8    4.5    2.1                                       17    (1.1)    1.6    3.6    4.1    2.4                                       18    17.7     9.1    2.9    2.5    2.7                                 r-Ag5       20.9     15.4   12.3   20.8   7.0                                 Blank       3970 cpm 5640 cpm                                                                             3130 cpm                                                                             1730 cpm                                                                             1050 cpm                            ______________________________________                                         NOTES                                                                         .sup.1 Dominant T cell epitopes are defined as those peptides which showe     stimulation indices of ≧4 in at least one of 5 strains of mice         tested.                                                                       .sup.2 Peptides in bold face represent those with stimulation indices of      ≧4 in 2 or more strains of mice.                                       .sup.3 Stimulation index values are taken from FIGS. 5, 9, 10, 11 and 12,     in which spleen cells were from mice immunized 5 times with rAg5. The         exceptions are those values in parentheses which are taken from FIG. 4 in     which spleen cells were from 4 times rAg5 immunized mice.                

T cell epitopes of hornet Ag5 and a mouse testis protein Tpx. Crossreactivity of these two proteins was studied in mice immunized withtheir fragments (FIGS. 15 and 16). Spleen cells from BALB/c miceinmnunized with r-fragment Tpx-N (residue 14-106) responded equally wellto stimulation with the immunogen or the r-fragment IN of hornet Ag5(residue 1-114). Similarly spleen cells from BALB/c mice immunized withr-fragment Tpx-C (residue 101-163) responded equally well to stimulationwith the immunogen or the r-fragment C of hornet Ag5 (residue 151-204).

Proliferation of Tpx-N or -C primed spleen cells was also tested onstimulation with synthetic peptides which correspond to sequencespresent in r-fragment IN and C of hornet Ag5. The data in FIG. 15 showhigh stimulation indices (greater than 6) for both r-fragments Tpx-N andfragment IN, only moderate stimulation indices (about 3) for peptides 7,8 and 9, and near baseline stimulation indices of about 2 or less forthe remaining peptides. The data in FIG. 16 show high stimulationindices (greater than 15) for Tpx-C, 9 for fragment C, 6 for peptide 18,and 2 or less for all other peptides.

The data in FIGS. 15 and 16 were obtained from mice after 4immunizations. Further studies after five immunizations are planned asthe data in FIGS. 7 to 10 indicate that stronger stimulation indices areobtained after longer immunization.

Discussion

The present Example defines indices of 4 or greater for proliferation ofAg5- specific cells as containing a T cell epitope of Ag5. With thisdefinition, the data in FIGS. 3-13 for mice of 5 strains with differenthaplotypes would indicate that the T cell epitopes of Ag5 aredistributed throughout the entire molecule. This is summarized in Table4. As C57B1/6 mice is a poor responder, its data are excluded in thefollowing comparison.

The data in Table 4 indicate that 14 peptides contain T cell epitopes ofAg5. Three of these peptides are recognized by three of the four strainstested, five of them by two strains and the remaining six by only onestrain. The peptides which are recognized by two or three strains ofmice are indicated in bold face characters in Table 2. They are peptides1, 2, 5, 6, 9, 15, 20 and 18. As can be seen in FIG. 1, there areextensive sequence identities of these white faced hornet peptides withthose of the other vespids. This is particularly apparent for peptides6, 20, and 18.

In addition to the seven vespid Ag5 sequences in FIG. 1, there are eightother vespid Ag5s with known sequences. They are nearly identical tothose in FIG. 1, depending on their species group [Lu, et al., J.Immunol., 150:2823-2830 (1993); Hoffman, J. Allergy Clin. Immunol.,92:707-716 (1993)]. Fire ant venom allergen Sol i 3 also has sequencesimilarity with vespid Ag5s [Hoffman, J. Allergy Clin. Immunol.,91:71-78 (1993)]. Peptides 9, 20 and 18 of white faced hornet Ag5 have ahigh degree of sequence identity with the corresponding peptides of Soli 3.

It is most likely that these vespid and fire ant Ag5s have T cellepitopes of identical and/or similar sequences. These epitopes studiesare being continued with yellowjacket or paper wasp Ag5 immunized mice.

The finding of cross reactive T cell epitopes of hornet Ag5 and amammalian testis protein is interesting, although its significance inclinical allergy, if any, is unknown. Published reports do not indicateany unusual distribution of male and female insect allergic patients.Our own unpublished studies showed that male and female BALB/c mice gaveindisguisable antibody response when immunized with hornet Ag5 inpresence of alum.

The present invention is not to be limited in scope by the specificembodiments describe herein. Indeed, various modifications of theinvention in addition to those described herein will become apparent tothose skilled in the art from the foregoing description and theaccompanying figures. Such modifications are intended to fall within thescope of the appended claims.

It is further to be understood that all base sizes or amino acid sizes,and all molecular weight or molecular mass values, given for nucleicacids or polypeptides are approximate, and are provided for description.

Various publications are cited herein, the disclosures of which areincorporated by reference in their entireties.

    __________________________________________________________________________    #             SEQUENCE LISTING                                                - (1) GENERAL INFORMATION:                                                    -    (iii) NUMBER OF SEQUENCES: 81                                            - (2) INFORMATION FOR SEQ ID NO:1:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 204 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Vespula m - #aculifrons                               -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                 - Asn Asn Tyr Cys Lys Ile Lys Cys Leu Lys Gl - #y Gly Val His Thr Ala         #                15                                                           - Cys Lys Tyr Gly Ser Leu Lys Pro Asn Cys Gl - #y Asn Lys Lys Val Val         #            30                                                               - Ser Tyr Gly Leu Thr Lys Gln Glu Lys Gln As - #p Ile Leu Lys Glu His         #        45                                                                   - Asn Asp Phe Arg Gln Lys Ile Ala Arg Gly Le - #u Glu Thr Arg Gly Asn         #    60                                                                       - Pro Gly Pro Gln Pro Pro Ala Lys Asn Met Ly - #s Asn Leu Val Trp Ser         #80                                                                           - Asp Glu Leu Ala Tyr Ile Ala Gln Val Trp Al - #a Asn Gln Cys Gln Tyr         #                95                                                           - Gly His Asp Thr Cys Arg Asp Val Ala Lys Ty - #r Gln Val Gly Gln Asn         #           110                                                               - Val Ala Leu Thr Gly Ser Thr Ala Ala Val Ty - #r Asn Asp Pro Val Lys         #       125                                                                   - Leu Val Lys Met Trp Glu Asp Glu Val Lys As - #p Tyr Asn Pro Lys Lys         #   140                                                                       - Lys Phe Ser Glu Asn Asn Phe Leu Lys Ile Gl - #y His Tyr Thr Gln Met         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Val Trp Ala Asn Thr Lys Glu Val Gly Cys Gl - #y Ser Ile Lys Tyr Ile         #               175                                                           - Gln Glu Asn Trp His Lys His Tyr Leu Val Cy - #s Asn Tyr Gly Pro Ser         #           190                                                               - Gly Asn Phe Gln Asn Glu Glu Leu Tyr Gln Th - #r Lys                         #       200                                                                   - (2) INFORMATION FOR SEQ ID NO:2:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 204 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Vespula v - #ulgaris                                  -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                 - Asn Asn Tyr Cys Lys Ile Lys Cys Leu Lys Gl - #y Gly Val His Thr Ala         #                15                                                           - Cys Lys Tyr Gly Ser Leu Lys Pro Asn Cys Gl - #y Asn Lys Val Val Val         #            30                                                               - Ser Tyr Gly Leu Thr Lys Gln Glu Lys Gln As - #p Ile Leu Lys Glu His         #        45                                                                   - Asn Asp Phe Arg Gln Lys Ile Ala Arg Gly Le - #u Glu Thr Arg Gly Asn         #    60                                                                       - Pro Gly Pro Gln Pro Pro Ala Lys Asn Met Ly - #s Asn Leu Val Trp Asn         #80                                                                           - Asp Glu Leu Ala Tyr Val Ala Gln Val Trp Al - #a Asn Gln Cys Gln Tyr         #                95                                                           - Gly His Asp Thr Cys Arg Asp Val Ala Lys Ty - #r Gln Val Gly Gln Asn         #           110                                                               - Val Ala Leu Thr Gly Ser Thr Ala Ala Lys Ty - #r Asp Asp Pro Val Lys         #       125                                                                   - Leu Val Lys Met Trp Glu Asp Glu Val Lys As - #p Tyr Asn Pro Lys Lys         #   140                                                                       - Lys Phe Ser Gly Asn Asp Phe Leu Lys Thr Gl - #y His Tyr Thr Gln Met         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Val Trp Ala Asn Thr Lys Glu Val Gly Cys Gl - #y Ser Ile Lys Tyr Ile         #               175                                                           - Gln Glu Lys Trp His Lys His Tyr Leu Val Cy - #s Asn Tyr Gly Pro Ser         #           190                                                               - Gly Asn Phe Met Asn Glu Glu Leu Tyr Gln Th - #r Lys                         #       200                                                                   - (2) INFORMATION FOR SEQ ID NO:3:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 203 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Dolichovespu - #la arenaria                           -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                 - Asn Asn Tyr Cys Lys Ile Cys Pro Lys Gly Th - #r His Thr Leu Cys Lys         #                15                                                           - Tyr Gly Thr Ser Met Lys Pro Asn Cys Gly Gl - #y Lys Ile Val Lys Ser         #            30                                                               - Tyr Gly Val Thr Asn Asp Glu Lys Asn Glu Il - #e Val Lys Arg His Asn         #        45                                                                   - Glu Phe Arg Gln Lys Val Ala Gln Gly Leu Gl - #u Thr Arg Gly Asn Pro         #    60                                                                       - Gly Pro Gln Pro Pro Ala Lys Asn Met Asn Le - #u Leu Val Trp Asn Asp         #80                                                                           - Glu Leu Ala Lys Ile Ala Gln Thr Trp Ala As - #n Gln Cys Asn Phe Gly         #                95                                                           - His Asp Gln Cys Arg Asn Thr Ala Lys Tyr Pr - #o Val Gly Gln Asn Val         #           110                                                               - Ala Ile Ala Ser Thr Thr Gly Asn Ser Tyr Gl - #n Thr Met Ser Tyr Leu         #       125                                                                   - Ile Lys Met Trp Glu Asp Glu Val Lys Asp Ty - #r Asn Pro His Lys Asp         #   140                                                                       - Leu Met His Asn Asn Phe Ser Lys Val Gly Hi - #s Tyr Thr Gln Met Val         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Trp Gly Lys Thr Lys Glu Ile Gly Cys Gly Se - #r Val Lys Tyr Ile Glu         #               175                                                           - Asn Lys Trp His Thr His Tyr Leu Val Cys As - #n Tyr Gly Pro Ala Gly         #           190                                                               - Asn Tyr Met Asn Gln Pro Val Tyr Glu Arg Ly - #s                             #       200                                                                   - (2) INFORMATION FOR SEQ ID NO:4:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 205 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Dolichovespu - #la maculata                           -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                 - Asn Asn Tyr Cys Lys Ile Lys Cys Ser Arg Gl - #y Ile His Thr Leu Cys         #                15                                                           - Lys Phe Gly Thr Ser Met Lys Pro Asn Cys Gl - #y Ser Lys Ile Val Lys         #            30                                                               - Val His Gly Val Ser Asn Asp Glu Lys Asn Gl - #u Ile Val Asn Arg His         #        45                                                                   - Asn Gln Phe Arg Gln Lys Val Ala Lys Gly Le - #u Glu Thr Arg Gly Asn         #    60                                                                       - Pro Gly Pro Gln Pro Pro Ala Lys Asn Met As - #n Val Leu Val Trp Asn         #80                                                                           - Asp Glu Leu Ala Lys Ile Ala Gln Thr Trp Al - #a Asn Gln Cys Ser Phe         #                95                                                           - Gly His Asp Gln Cys Arg Asn Thr Glu Lys Ty - #r Gln Val Gly Gln Asn         #           110                                                               - Val Ala Ile Ala Ser Thr Thr Gly Asn Ser Ty - #r Ala Thr Met Ser Lys         #       125                                                                   - Leu Ile Glu Met Trp Glu Asn Glu Val Lys As - #p Phe Asn Pro Lys Lys         #   140                                                                       - Gly Thr Ile Gly Asp Asn Asn Phe Ser Lys Va - #l Gly His Tyr Thr Gln         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Met Val Trp Gly Lys Thr Lys Glu Ile Gly Cy - #s Gly Ser Val Lys Tyr         #               175                                                           - Ile Glu Asn Asn Trp His Thr His Tyr Leu Va - #l Cys Asn Tyr Gly Pro         #           190                                                               - Ala Gly Asn Tyr Met Asp Gln Pro Ile Tyr Gl - #u Arg Lys                     #       205                                                                   - (2) INFORMATION FOR SEQ ID NO:5:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 204 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:                                                     -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Dolichovespu - #la maculata                           -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                 - Asn Asn Tyr Cys Lys Ile Lys Cys Arg Lys Gl - #y Ile His Thr Leu Cys         #                15                                                           - Lys Phe Gly Thr Ser Met Lys Pro Asn Cys Gl - #y Arg Asn Val Val Lys         #            30                                                               - Ala Tyr Gly Leu Thr Asn Asp Glu Lys Asn Gl - #u Ile Leu Lys Arg His         #        45                                                                   - Asn Asp Phe Arg Gln Asn Val Ala Lys Gly Le - #u Glu Thr Arg Gly Lys         #    60                                                                       - Pro Gly Pro Gln Pro Pro Ala Lys Asn Met As - #n Val Leu Val Trp Asn         #80                                                                           - Asp Glu Leu Ala Lys Ile Ala Gln Thr Trp Al - #a Asn Gln Cys Asp Phe         #                95                                                           - Asn His Asp Asp Cys Arg Asn Thr Ala Lys Ty - #r Gln Val Gly Gln Asn         #           110                                                               - Ile Ala Ile Ser Ser Thr Thr Ala Thr Gln Ph - #e Asp Arg Pro Ser Lys         #       125                                                                   - Leu Ile Lys Gln Trp Glu Asp Glu Val Thr Gl - #u Phe Asn Tyr Lys Val         #   140                                                                       - Gly Leu Gln Asn Ser Asn Phe Arg Lys Val Gl - #y His Tyr Thr Gln Met         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Val Trp Gly Lys Thr Lys Glu Ile Gly Cys Gl - #y Ser Ile Lys Tyr Ile         #               175                                                           - Glu Asp Asn Trp Tyr Thr His Tyr Leu Val Cy - #s Asn Tyr Gly Pro Gly         #           190                                                               - Gly Asn Asp Phe Asn Gln Pro Ile Tyr Glu Ar - #g Lys                         #       200                                                                   - (2) INFORMATION FOR SEQ ID NO:6:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 205 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:                                                     -     (vi) ORIGINAL SOURCE:                                                   #annularis(A) ORGANISM: Polistes                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                 - Val Asp Tyr Cys Lys Ile Lys Cys Pro Ser Gl - #y Ile His Thr Val Cys         #                15                                                           - Gln Tyr Gly Glu Ser Thr Lys Pro Ser Lys As - #n Cys Ala Gly Lys Val         #            30                                                               - Ile Lys Ser Val Gly Pro Thr Glu Glu Glu Ly - #s Lys Leu Ile Val Ser         #        45                                                                   - Glu His Asn Arg Phe Arg Gln Lys Val Ala Gl - #n Gly Leu Glu Thr Arg         #    60                                                                       - Gly Asn Pro Gly Pro Gln Pro Ala Ala Ser As - #p Met Asn Asp Leu Val         #80                                                                           - Trp Asn Asp Glu Leu Ala His Ile Ala Gln Va - #l Trp Ala Ser Gln Cys         #                95                                                           - Gln Phe Leu Val His Asp Lys Cys Arg Asn Th - #r Ala Lys Tyr Pro Val         #           110                                                               - Gly Gln Asn Ile Ala Tyr Ala Gly Gly Ser As - #n Leu Pro Asp Val Val         #       125                                                                   - Ser Leu Ile Lys Leu Trp Glu Asn Glu Val Ly - #s Asp Phe Asn Tyr Asn         #   140                                                                       - Thr Gly Ile Thr Lys Gln Asn Phe Ala Lys Il - #e Gly His Tyr Thr Gln         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Met Val Trp Gly Lys Thr Lys Glu Ile Gly Cy - #s Gly Ser Leu Lys Tyr         #               175                                                           - Met Glu Asn Asn Met Gln Asn His Tyr Leu Il - #e Cys Asn Tyr Gly Pro         #           190                                                               - Ala Gly Asn Tyr Leu Gly Gln Leu Pro Tyr Th - #r Lys Lys                     #       205                                                                   - (2) INFORMATION FOR SEQ ID NO:7:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 205 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:                                                     -     (vi) ORIGINAL SOURCE:                                                   #exclamans(A) ORGANISM: Polistes                                              -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                 - Val Asp Tyr Cys Lys Ile Lys Cys Pro Ser Gl - #y Ile His Thr Val Cys         #                15                                                           - Gln Tyr Gly Glu Ser Thr Lys Pro Ser Lys As - #n Cys Ala Gly Lys Val         #            30                                                               - Ile Lys Ser Val Gly Pro Thr Glu Glu Glu Ly - #s Lys Leu Ile Val Ser         #        45                                                                   - Glu His Asn Arg Phe Arg Gln Lys Val Ala Gl - #n Gly Leu Glu Thr Arg         #    60                                                                       - Gly Asn Pro Gly Pro Gln Pro Ala Ala Ser As - #p Met Asn Asp Leu Val         #80                                                                           - Trp Asn Asp Glu Leu Ala His Ile Ala Gln Va - #l Trp Ala Ser Gln Cys         #                95                                                           - Gln Phe Leu Val His Asp Lys Cys Arg Asn Th - #r Ala Lys Tyr Pro Val         #           110                                                               - Gly Gln Asn Ile Ala Tyr Ala Gly Gly Ser Ly - #s Leu Pro Asp Val Val         #       125                                                                   - Ser Leu Ile Lys Leu Trp Glu Asn Glu Val Ly - #s Asp Phe Asn Tyr Asn         #   140                                                                       - Thr Gly Ile Thr Lys Gln Asn Phe Ala Lys Il - #e Gly His Tyr Thr Gln         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Met Val Trp Gly Lys Thr Lys Glu Ile Gly Cy - #s Gly Ser Leu Lys Tyr         #               175                                                           - Ile Glu Asn Lys Met Gln Asn His Tyr Leu Il - #e Cys Asn Tyr Gly Pro         #           190                                                               - Ala Gly Asn Tyr Leu Gly Gln Leu Pro Tyr Th - #r Lys Lys                     #       205                                                                   - (2) INFORMATION FOR SEQ ID NO:8:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:  N-terminal                                         -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                 - Asn Asn Tyr Cys Lys Ile Lys Cys Arg Lys Gl - #y Ile His Thr Leu Cys         #                15                                                           - Lys Phe Gly Thr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:9:                                            -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                 - Gly Ile His Thr Leu Cys Lys Phe Gly Thr Se - #r Met Lys Pro Asn Cys         #                15                                                           - Gly Arg Asn Val                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:10:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                - Ser Met Lys Pro Asn Cys Gly Arg Asn Val Va - #l Lys Ala Tyr Gly Leu         #                15                                                           - Thr Asn Asp Glu                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:11:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                - Val Lys Ala Tyr Gly Leu Thr Asn Asp Glu Ly - #s Asn Glu Ile Leu Lys         #                15                                                           - Arg His Asn Asp                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:12:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                - Lys Asn Glu Ile Leu Lys Arg His Asn Asp Ph - #e Arg Gln Asn Val Ala         #                15                                                           - Lys Gly Leu Glu                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:13:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                - Phe Arg Gln Asn Val Ala Lys Gly Leu Glu Th - #r Arg Gly Lys Pro Gly         #                15                                                           - Pro Gln Pro Pro                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:14:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                - Thr Arg Gly Lys Pro Gly Pro Gln Pro Pro Al - #a Lys Asn Met Asn Val         #                15                                                           - Leu Val Trp Asn                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:15:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                - Ala Lys Asn Met Asn Val Leu Val Trp Asn As - #p Glu Leu Ala Lys Ile         #                15                                                           - Ala Gln Thr Trp                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:16:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                - Asp Glu Leu Ala Lys Ile Ala Gln Thr Trp Al - #a Asn Gln Cys Asp Phe         #                15                                                           - Asn His Asp Asp                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:17:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                - Ala Asn Gln Cys Asp Phe Asn His Asp Asp Cy - #s Arg Asn Thr Ala Lys         #                15                                                           - Tyr Gln Val Gly                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:18:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                - Cys Arg Asn Thr Ala Lys Tyr Gln Val Gly Gl - #n Asn Ile Ala Ile Ser         #                15                                                           - Ser Thr Thr Ala                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:19:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                - Gln Asn Ile Ala Ile Ser Ser Thr Thr Ala Th - #r Gln Phe Asp Arg Pro         #                15                                                           - Ser Lys Leu Ile                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:20:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                - Thr Gln Phe Asp Arg Pro Ser Lys Leu Ile Ly - #s Gln Trp Glu Asp Glu         #                15                                                           - Val Thr Glu Phe                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:21:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                - Lys Gln Trp Glu Asp Glu Val Thr Glu Phe As - #n Tyr Lys Val Gly Leu         #                15                                                           - Gln Asn Ser Asn                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:22:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                - Asn Tyr Lys Val Gly Leu Gln Asn Ser Asn Ph - #e Arg Lys Val Gly His         #                15                                                           - Tyr Thr Gln Met                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:23:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                - Phe Arg Lys Val Gly His Tyr Thr Gln Met Va - #l Trp Gly Lys Thr             #                15                                                           - NFORMATION FOR SEQ ID NO:24:                                                -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                - His Tyr Thr Gln Met Val Trp Gly Lys Thr Ly - #s Glu Ile Gly Cys Gly         #                15                                                           - Ser Ile Lys Tyr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:25:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                - Lys Glu Ile Gly Cys Gly Ser Ile Lys Tyr Il - #e Glu Asp Asn Trp Tyr         #                15                                                           - Thr His Tyr Leu                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:26:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                - Ile Glu Asp Asn Trp Tyr Thr His Tyr Leu Va - #l Cys Asn Tyr Gly Pro         #                15                                                           - Gly Gly Asn Asp                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:27:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 19 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE:  C-terminal                                         -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                - Val Cys Asn Tyr Gly Pro Gly Gly Asn Asp Ph - #e Asn Gln Pro Ile Tyr         #                15                                                           - Glu Arg Lys                                                                 - (2) INFORMATION FOR SEQ ID NO:28:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 151 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: N-terminal                                          -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Homo sapi - #ens                                      -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                - Gln Val Gln Arg Glu Ile Val Asn Lys His As - #n Glu Leu Arg Lys Ala         #                15                                                           - Val Ser Pro Pro Ala Ser Asn Met Leu Lys Me - #t Glu Trp Ser Arg Glu         #            30                                                               - Val Thr Thr Asn Ala Gln Arg Trp Ala Asn Ly - #s Cys Thr Leu Gln His         #        45                                                                   - Ser Asp Pro Glu Asp Arg Lys Thr Ser Thr Ar - #g Cys Gly Glu Asn Leu         #    60                                                                       - Tyr Met Ser Ser Asp Pro Thr Ser Trp Ser Se - #r Ala Ile Gln Ser Trp         #80                                                                           - Tyr Asp Glu Ile Leu Asp Phe Val Tyr Gly Va - #l Gly Pro Lys Ser Pro         #                95                                                           - Asn Ala Val Val Gly His Tyr Thr Gln Leu Va - #l Trp Tyr Ser Thr Tyr         #           110                                                               - Gln Val Gly Cys Gly Ile Ala Tyr Cys Pro As - #n Gln Asp Ser Leu Lys         #       125                                                                   - Tyr Tyr Tyr Val Cys Gln Tyr Cys Pro Ala Gl - #y Asn Asn Met Asn Arg         #   140                                                                       - Lys Asn Thr Pro Tyr Gln Gln                                                 145                 1 - #50                                                   - (2) INFORMATION FOR SEQ ID NO:29:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 150 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: N-terminal                                          -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Mus muscu - #lus                                      -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                - Gln Val Gln Arg Glu Ile Val Asn Lys His As - #n Glu Leu Arg Arg Ser         #                15                                                           - Val Asn Pro Thr Gly Ser Asp Ile Leu Lys Me - #t Glu Trp Ser Ile Gln         #            30                                                               - Ala Thr Thr Asn Ala Gln Lys Trp Ala Asn Ly - #s Cys Ile Leu Glu His         #        45                                                                   - Ser Ser Lys Asp Asp Arg Lys Ile Asn Ile Ar - #g Cys Gly Glu Asn Leu         #    60                                                                       - Tyr Met Ser Thr Asp Pro Thr Leu Trp Ser Th - #r Val Ile Gln Ser Trp         #80                                                                           - Tyr Asn Glu Asn Glu Asp Phe Val Tyr Gly Va - #l Gly Ala Lys Pro Asn         #                95                                                           - Ser Ala Val Gly His Tyr Thr Gln Leu Val Tr - #p Tyr Ser Ser Phe Lys         #           110                                                               - Ile Gly Cys Gly Ile Ala Tyr Cys Pro Asn Gl - #n Asp Asn Leu Lys Tyr         #       125                                                                   - Phe Tyr Val Cys His Tyr Cys Pro Met Gly As - #n Asn Val Met Lys Lys         #   140                                                                       - Ser Thr Pro Tyr Gln Gln                                                     145                 1 - #50                                                   - (2) INFORMATION FOR SEQ ID NO:30:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 166 amino                                                         (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: protein                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: N-terminal                                          -     (vi) ORIGINAL SOURCE:                                                             (A) ORGANISM: Vespid                                                -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                - Asp Glu Lys Asn Glu Ile Leu Lys Arg His As - #n Asp Phe Arg Gln Asn         #                15                                                           - Val Ala Lys Gly Leu Glu Thr Arg Gly Lys Pr - #o Gly Pro Gln Pro Pro         #            30                                                               - Ala Lys Asn Met Asn Val Leu Val Trp Asn As - #p Glu Leu Ala Lys Ile         #        45                                                                   - Ala Gln Thr Trp Ala Asn Gln Cys Asp Phe As - #n His Asp Asp Cys Arg         #    60                                                                       - Asn Thr Ala Lys Tyr Gln Val Gly Gln Asn Il - #e Ala Ile Ser Ser Thr         #80                                                                           - Thr Ala Thr Gln Phe Asp Arg Pro Ser Lys Le - #u Ile Lys Gln Trp Glu         #                95                                                           - Asp Glu Val Thr Glu Phe Asn Tyr Lys Val Gl - #y Leu Gln Asn Ser Asn         #           110                                                               - Phe Arg Lys Val Gly His Tyr Thr Gln Met Va - #l Trp Gly Lys Thr Lys         #       125                                                                   - Glu Ile Gly Cys Gly Ser Ile Lys Tyr Ile Gl - #u Asp Asn Trp Tyr Thr         #   140                                                                       - His Tyr Leu Val Cys Asn Tyr Gly Pro Gly Gl - #y Asn Asp Phe Asn Gln         145                 1 - #50                 1 - #55                 1 -       #60                                                                           - Pro Ile Tyr Glu Arg Lys                                                                     165                                                           - (2) INFORMATION FOR SEQ ID NO:31:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: YES                                                  -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                - Xaa Asx Tyr Cys Lys Ile Xaa Cys Xaa Xaa Gl - #y Xaa Xaa His Thr Xaa         #                15                                                           - Cys Xaa Xaa Gly                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:32:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 24 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: YES                                                  -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                - Gly Xaa Xaa His Thr Xaa Cys Xaa Xaa Gly Xa - #a Ser Xaa Lys Pro Xaa         #                15                                                           - Xaa Asn Cys Xaa Xaa Xaa Xaa Xaa                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:33:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 34 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: YES                                                  -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                - Xaa Asx Tyr Cys Lys Ile Xaa Cys Xaa Xaa Gl - #y Xaa Xaa His Thr Xaa         #                15                                                           - Cys Xaa Xaa Gly Xaa Ser Xaa Lys Pro Xaa Xa - #a Asn Cys Xaa Xaa Xaa         #            30                                                               - Xaa Xaa                                                                     - (2) INFORMATION FOR SEQ ID NO:34:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                - Lys Xaa Xaa Ile Xaa Xaa Xaa His Asn Xaa Ph - #e Arg Gln Lys Xaa Ala         #                15                                                           - Xaa Gly Leu Glu                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:35:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                - Phe Arg Gln Lys Xaa Ala Xaa Gly Leu Glu Th - #r Arg Gly Xaa Pro Gly         #                15                                                           - Pro Gln Pro Xaa                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:36:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 30 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                - Lys Xaa Xaa Ile Xaa Xaa Xaa His Asn Xaa Ph - #e Arg Gln Lys Xaa Ala         #                15                                                           - Xaa Gly Leu Glu Thr Arg Gly Xaa Pro Gly Pr - #o Gln Pro Xaa                 #            30                                                               - (2) INFORMATION FOR SEQ ID NO:37:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                - Asp Glu Leu Ala Xaa Xaa Ala Gln Xaa Trp Al - #a Xaa Gln Cys Xaa Xaa         #                15                                                           - Xaa Xaa His Asp                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:38:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                - Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asx Xa - #a Phe Xaa Lys Xaa Gly         #                15                                                           - His Tyr Thr Gln Met                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:39:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 15 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                - Phe Xaa Lys Xaa Gly His Tyr Thr Gln Met Va - #l Trp Xaa Xaa Thr             #                15                                                           - NFORMATION FOR SEQ ID NO:40:                                                -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 26 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                - Asn Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asx Xa - #a Phe Xaa Lys Xaa Gly         #                15                                                           - His Tyr Thr Gln Met Val Trp Xaa Xaa Thr                                     #            25                                                               - (2) INFORMATION FOR SEQ ID NO:41:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 22 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                - Xaa Glx Xaa Xaa Xaa Xaa Xaa His Tyr Leu Xa - #a Cys Asn Tyr Gly Pro         #                15                                                           - Xaa Gly Asn Xaa Xaa Xaa                                                                 20                                                                - (2) INFORMATION FOR SEQ ID NO:42:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 32 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                - Asn Asn Tyr Cys Lys Ile Lys Cys Arg Lys Gl - #y Ile His Thr Leu Cys         #                15                                                           - Lys Phe Gly Thr Gly Thr Ser Met Lys Pro As - #n Cys Gly Arg Asn Val         #            30                                                               - (2) INFORMATION FOR SEQ ID NO:43:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 30 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                - Lys Asn Glu Ile Leu Lys Arg His Asn Asp Ph - #e Arg Gln Asn Val Ala         #                15                                                           - Lys Gly Leu Glu Thr Arg Gly Lys Pro Gly Pr - #o Gln Pro Pro                 #            30                                                               - (2) INFORMATION FOR SEQ ID NO:44:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 25 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                - Asn Tyr Lys Val Gly Leu Gln Asn Ser Asn Ph - #e Arg Lys Val Gly His         #                15                                                           - Tyr Thr Gln Met Val Trp Gly Lys Thr                                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:45:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                - Asn Asn Tyr Cys Lys Ile Lys Cys Leu Lys Gl - #y Gly Val His Thr Ala         #                15                                                           - Cys Lys Tyr Gly Ser Leu Lys Pro Asn Cys Gl - #y Asn Lys Lys Val             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:46:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                - Asn Asn Tyr Cys Lys Ile Lys Cys Leu Lys Gl - #y Gly Val His Thr Ala         #                15                                                           - Cys Lys Tyr Gly Ser Leu Lys Pro Asn Cys Gl - #y Asn Lys Val Val             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:47:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                - Asn Asn Tyr Cys Lys Ile Cys Pro Lys Gly Th - #r His Thr Leu Cys Lys         #                15                                                           - Tyr Gly Thr Ser Met Lys Pro Asn Cys Gly Gl - #y Lys Ile Val Lys             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:48:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 32 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                - Asn Asn Tyr Cys Lys Ile Lys Cys Ser Arg Gl - #y Ile His Thr Leu Cys         #                15                                                           - Lys Phe Gly Thr Ser Met Lys Pro Asn Cys Gl - #y Ser Lys Ile Val Lys         #            30                                                               - (2) INFORMATION FOR SEQ ID NO:49:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 32 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                - Asn Asn Tyr Cys Lys Ile Lys Cys Arg Lys Gl - #y Ile His Thr Leu Cys         #                15                                                           - Lys Phe Gly Thr Ser Met Lys Pro Asn Cys Gl - #y Arg Asn Val Val Lys         #            30                                                               - (2) INFORMATION FOR SEQ ID NO:50:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 34 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                - Val Asp Tyr Cys Lys Ile Lys Cys Pro Ser Gl - #y Ile His Thr Val Cys         #                15                                                           - Gln Tyr Gly Glu Ser Thr Lys Pro Ser Lys As - #n Cys Ala Gly Lys Val         #            30                                                               -      Ile Lys                                                                - (2) INFORMATION FOR SEQ ID NO:51:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 34 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                - Val Asp Tyr Cys Lys Ile Lys Cys Pro Ser Gl - #y Ile His Thr Val Cys         #                15                                                           - Gln Tyr Gly Glu Ser Thr Lys Pro Ser Lys As - #n Cys Ala Gly Lys Val         #            30                                                               - Ile Lys                                                                     - (2) INFORMATION FOR SEQ ID NO:52:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 35 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                - Tyr Asn Tyr Cys Asn Leu Gln Ser Cys Lys Ar - #g Asn Asn Ala Ile His         #                15                                                           - Thr Met Cys Gln Tyr Thr Ser Pro Thr Pro Gl - #y Pro Met Cys Leu Glu         #            30                                                               - Tyr Ser Asn                                                                         35                                                                    - (2) INFORMATION FOR SEQ ID NO:53:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                - Lys Gln Asp Ile Leu Lys Glu His Asn Asp Ph - #e Arg Gln Lys Ile Ala         #                15                                                           - Arg Gly Leu Glu Thr Arg Gly Asn Pro Gly Pr - #o Gln Pro Pro Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:54:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                - Lys Gln Asp Ile Leu Lys Glu His Asn Asp Ph - #e Arg Gln Lys Ile Ala         #                15                                                           - Arg Gly Leu Glu Thr Arg Gly Asn Pro Gly Pr - #o Gln Pro Pro Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:55:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                - Lys Asn Glu Ile Val Lys Arg His Asn Glu Ph - #e Arg Gln Lys Val Ala         #                15                                                           - Gln Gly Leu Glu Thr Arg Gly Asn Pro Gly Pr - #o Gln Pro Pro Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:56:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                - Lys Asn Glu Ile Val Asn Arg His Asn Gln Ph - #e Arg Gln Lys Val Ala         #                15                                                           - Lys Gly Leu Glu Thr Arg Gly Asn Pro Gly Pr - #o Gln Pro Pro Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:57:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                - Lys Asn Glu Ile Leu Lys Arg His Asn Asp Ph - #e Arg Gln Asn Val Ala         #                15                                                           - Lys Gly Leu Glu Thr Arg Gly Lys Pro Gly Pr - #o Gln Pro Pro Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:58:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                - Lys Lys Leu Ile Val Ser Glu His Asn Arg Ph - #e Arg Gln Lys Val Ala         #                15                                                           - Gln Gly Leu Glu Thr Arg Gly Asn Pro Gly Pr - #o Gln Pro Ala Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:59:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                - Lys Lys Leu Ile Val Ser Glu His Asn Arg Ph - #e Arg Gln Lys Val Ala         #                15                                                           - Gln Gly Leu Glu Thr Arg Gly Asn Pro Gly Pr - #o Gln Pro Ala Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:60:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 31 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                - Lys Asp Ala Ile Val Asn Lys His Asn Glu Le - #u Arg Gln Arg Val Ala         #                15                                                           - Ser Gly Lys Glu Met Arg Gly Thr Asn Gly Pr - #o Gln Pro Pro Ala             #            30                                                               - (2) INFORMATION FOR SEQ ID NO:61:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                - Asp Glu Leu Ala Tyr Ile Ala Gln Val Trp Al - #a Asn Gln Cys Gln Tyr         #                15                                                           - Gly His Asp Thr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:62:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                - Asp Glu Leu Ala Tyr Val Ala Gln Val Trp Al - #a Asn Gln Cys Gln Tyr         #                15                                                           - Gly His Asp Thr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:63:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                - Asp Glu Leu Ala Lys Ile Ala Gln Thr Trp Al - #a Asn Gln Cys Asn Phe         #                15                                                           - Gly His Asp Gln                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:64:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                - Asp Glu Leu Ala Lys Ile Ala Gln Thr Trp Al - #a Asn Gln Cys Ser Phe         #                15                                                           - Gly His Asp Gln                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:65:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                - Asp Glu Leu Ala His Ile Ala Gln Val Trp Al - #a Ser Gln Cys Gln Phe         #                15                                                           - Leu Val His Asp Lys                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:66:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 21 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                - Asp Glu Leu Ala His Ile Ala Gln Val Trp Al - #a Ser Gln Cys Gln Phe         #                15                                                           - Leu Val His Asp Lys                                                                     20                                                                - (2) INFORMATION FOR SEQ ID NO:67:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                - Pro Glu Leu Ala Thr Ile Ala Gln Arg Trp Al - #a Asn Gln Cys Thr Glu         #                15                                                           - Glu His Asp Ala                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:68:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 25 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                - Asn Pro Lys Lys Lys Phe Ser Glu Asn Asn Ph - #e Leu Lys Ile Gly His         #                15                                                           - Tyr Thr Gln Met Val Trp Ala Asn Thr                                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:69:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 25 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                - Asn Pro Lys Lys Lys Phe Ser Gly Asn Asp Ph - #e Leu Lys Thr Gly His         #                15                                                           - Tyr Thr Gln Met Val Trp Ala Asn Thr                                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:70:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 25 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                - Asn Pro His Lys Asp Leu Met His Asn Asn Ph - #e Ser Lys Val Gly His         #                15                                                           - Tyr Thr Gln Met Val Trp Gly Lys Thr                                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:71:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 26 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                - Asn Pro Lys Lys Gly Thr Ile Gly Asp Asn As - #n Phe Ser Lys Val Gly         #                15                                                           - His Tyr Thr Gln Met Val Trp Gly Lys Thr                                     #            25                                                               - (2) INFORMATION FOR SEQ ID NO:72:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 25 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                - Asn Tyr Asn Thr Gly Ile Thr Lys Gln Asn Ph - #e Ala Lys Ile Gly His         #                15                                                           - Tyr Thr Gln Met Val Trp Gly Lys Thr                                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:73:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 25 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                - Asn Tyr Asn Thr Gly Ile Thr Lys Gln Asn Ph - #e Ala Lys Ile Gly His         #                15                                                           - Tyr Thr Gln Met Val Trp Gly Lys Thr                                         #            25                                                               - (2) INFORMATION FOR SEQ ID NO:74:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 29 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                - Asn Tyr Asn Thr Gly Ile Ser Phe Pro Ser As - #p Asp Asn Ile Leu Met         #                15                                                           - Lys Val Glu His Tyr Thr Gln Ile Val Trp Al - #a Lys Thr                     #            25                                                               - (2) INFORMATION FOR SEQ ID NO:75:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                - Ile Gln Glu Asn Trp His Lys His Tyr Leu Va - #l Cys Asn Tyr Gly Pro         #                15                                                           - Ser Gly Asn Phe                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:76:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                - Ile Gln Glu Lys Trp His Lys His Tyr Leu Va - #l Cys Asn Tyr Gly Pro         #                15                                                           - Ser Gly Asn Phe                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:77:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                - Ile Glu Asn Lys Trp His Thr His Tyr Leu Va - #l Cys Asn Tyr Gly Pro         #                15                                                           - Ala Gly Asn Tyr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:78:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                - Ile Glu Asn Asn Trp His Thr His Tyr Leu Va - #l Cys Asn Tyr Gly Pro         #                15                                                           - Ala Gly Asn Tyr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:79:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                - Met Glu Asn Asn Met Gln Asn His Tyr Leu Il - #e Cys Asn Tyr Gly Pro         #                15                                                           - Ala Gly Asn Tyr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:80:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                - Ile Glu Asn Lys Met Gln Asn His Tyr Leu Il - #e Cys Asn Tyr Gly Pro         #                15                                                           - Ala Gly Asn Tyr                                                                         20                                                                - (2) INFORMATION FOR SEQ ID NO:81:                                           -      (i) SEQUENCE CHARACTERISTICS:                                          #acids    (A) LENGTH: 20 amino                                                          (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                -     (ii) MOLECULE TYPE: peptide                                             -    (iii) HYPOTHETICAL: NO                                                   -     (iv) ANTI-SENSE: NO                                                     -      (v) FRAGMENT TYPE: internal                                            -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                - Glu Pro Asp Asn Trp Thr Lys His Tyr Leu Va - #l Cys Asn Tyr Gly Pro         #                15                                                           - Ala Gly Asn Val                                                                         20                                                                __________________________________________________________________________

What is claimed is:
 1. A method for treating sensitivity to vespid venomantigen 5 comprising administering to a vespid venom allergic patient atherapeutically effective amount of a peptide characterized by thefollowing properties:a) it has a range of about 8 to about 35 amino acidresidues of vespid venom antigen 5; and, b) it is antigenic for T cellproliferation in a mouse immunized with a vespid venom antigen 5, whichmouse is a strain selected from the group consisting of BALB/c, ASW/Sn,C3H/He, and P/J.
 2. The method according to claim 1, wherein the peptidecorresponds to a fragment of white face hornet antigen 5, form 2, havingan amino acid sequence selected form the group consisting of:

    NNYCKIKCRKGIHTLCKFGT; (SEQ ID NO:8)                                           GIHTLCKFGTSMKPNCGRNV; (SEQ ID NO:9)                                           KNEILKRHNDFRQNVAKGLE; (SEQ ID NO:12)                                          FRQNVAKGLETRGKPGPQPP; (SEQ ID NO:13)                                          TRGKPGPQPPAKNMNVLVWN; (SEQ ID NO:14)                                          DELAKIAQTWANQCDFNHDD; (SEQ ID NO:16)                                          CRNTAKYQVGQNIAISSTTA; (SEQ ID NO:18)                                          QNIAISSTTATQFDRPSKLI; (SEQ ID NO:19)                                          TQFDRPSKLIKQWEDEVTEF; (SEQ ID NO:20)                                          KQWEDEVTEFNYKVGLQNSN; (SEQ ID NO:21)                                          NYKVGLQNSNFRKVGHYTQM; (SEQ ID NO:22)                                          FRKVGHYTQMVWGKT;      (SEQ ID NO:23)                                          KEIGCGSIKYIEDNWYTHYL; and                                                                           (SEQ ID NO:25)                                          IEDNWYTHYLVCNYGPGGND. (SEQ ID NO:26)                                      


3. The method according to claim 1, wherein the peptide has a sequenceselected from the group consisting of:

    NNYCKIKCRKGIHTLCKFGT;                                                                             (SEQ ID NO:8)                                             GIHTLCKFGTSMKPNCGRNV;                                                                             (SEQ ID NO:9)                                             KNEILKRHNDFRQNVAKGLE;                                                                             (SEQ ID NO:12)                                            FRQNVAKGLETRGKPGPQPP;                                                                             (SEQ ID NO:13)                                            DELAKIAQTWANQCDFNHDD;                                                                             (SEQ ID NO:16)                                            NYKVGLQNSNFRKVGHYTQM;                                                                             (SEQ ID NO:22)                                            FRKVGHYTQMVWGKT; and                                                                              (SEQ ID NO:23)                                            IEDNWYTHYLVCNYGPGGND.                                                                             (SEQ ID NO:26)                                        


4. The method according to claim 1, wherein the patient has beenidentified as sensitive to vespid venom antigen
 5. 5. The methodaccording to claim 1, wherein the peptide is formulated in apharmaceutical composition.
 6. A method for treating sensitivity tovespid venom antigen 5 comprising administering to a vespid venomallergic patient a therapeutically effective amount of a peptide havingan amino acid sequence selected from the group consisting of:a)NNYCKIKCRKGIHTLCKFGT (SEQ ID NO:8), or its homolog; b)GIHTLCKFGTSMKPNCGRNV (SEQ ID NO:9), or its homolog; c)NNYCKIKCRKGIHTLCKFGTGTSMKPNCGRNV (SEQ ID NO:42), or its homolog; d)KNEILKRHNDFRQNVAKGLE (SEQ ID NO: 12), or its homolog; e)FRQNVAKGLETRGKPGPQPP (SEQ ID NO: 13), or its homolog; f)KNEILKRHNDFRQNVAKGLETRGKPGPQPP (SEQ ID NO:43), or its homolog; g)DELAKIAQTWANQCDFNHDD (SEQ ID NO:16), or its homolog; h)NYKVGLQNSNFRKVGHYTQM (SEQ ID NO:22), or its homolog; i) FRKVGHYTQMVWGKT(SEQ ID NO.23), or its homolog; j) NYKVGLQNSNFRKVGHYTQMVWGKT (SEQ IDNO:44), or its homolog; k) IEDNWYTHYLVCNYGPGGND (SEQ ID NO:26), or itshomolog.
 7. The method according, to claim 6, wherein the patient hasbeen identified as sensitive to vespid venom antigen
 5. 8. The methodaccording to claim 6, wherein the peptide is formulated in apharmaceutical composition.
 9. A method for treating sensitivity tovespid venom antigen 5 comprising administering to a vespid venomallergic patient a therapeutically effective amount of a recombinantpolypeptide comprising two or more peptides non-contiguously arrangedrelative to the native sequence of vespid venom antigen 5, wherein thepeptides are selected from the group consisting of:a)NNYCKIKRKGIHTLCKFGT (SEQ ID NO:8), or its homolog; b)GIHTLCKFGTSMKPNCGRNV (SEQ ID NO:9), or its homolog; c)NNYCKIKCRKGIHTLCKFGTGTSMKPNCGRNV (SEQ ID NO:42), or its homolog; d)KNEILKRHNDFRQNVAKGLE (SEQ ID NO:12), or its homolog; e)FRQNVAKGLETRGKPGPQPP (SEQ ID NO:13), or its homolog; f)KNEILKRHNDFRQNVAKGLETRGKPGPQPP (SEQ ID NO:43), or its homolog; g)DELAKIAQTWANQCDFNHDD (SEQ ID NO:16), or its homolog; h)NYKVGLQNSNFRKVGHYTQM (SEQ ID NO:22), or its homolog; i) FRKVGHYTQMVWGKT(SEQ ID NO:23), or its homolog; i) NYKVGLQNSNFRKVGHYTQMVWGKT (SEQ IDNO:44), or its homolog; and, k) IEDNWYTHYLVCNYGPGGND (SEQ ID NO:26), orits homolog.
 10. The method according to claim 9, wherein the patienthas been identified as sensitive to vespid venom antigen
 5. 11. Themethod according to claim 9, wherein the peptide is formulated in apharmaceutical composition.